KR20210091032A - Long acting conjugate of Trigonal glucagon/GLP-1/GIP receptor agonist for use in the treatment of pulmonary disease - Google Patents

Abstract

The present invention relates to a preventive or therapeutic use of a triple activator and/or a conjugate thereof against lung disease, the triple activator and/or conjugate thereof having activity with respect to all of glucagon and glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) receptors. The triple activator or long-acting conjugate thereof according to the present invention has activity on glucagon receptors, GLP-1 receptors, and GIP receptors, thereby being able to exhibit a preventive or therapeutic on lung diseases.

Description

Long acting conjugate of Trigonal glucagon/GLP-1/GIP receptor agonist for use in the treatment of pulmonary disease

The present invention relates to the prophylactic or therapeutic use of a triple activator and/or a conjugate thereof having activity on both glucagon, GLP-1 and GIP receptors for lung disease.

The lung is an organ mainly responsible for respiration, and lung diseases occur due to harmful substances, viruses, immune abnormalities, and the like. Since lung disease causes decreased lung function and respiratory discomfort, appropriate treatment is required according to the cause of the disease.

Lung-related diseases include interstitial lung disease, progressive fibrotic interstitial lung disease, idiopathic interstitial pneumonia, non-specific interstitial pneumonia, pulmonary fibrosis, interstitial pulmonary fibrosis, idiopathic pulmonary fibrosis, alveolitis, pneumonia, emphysema, bronchitis, chronic obstructive pulmonary disease (COPD), complex pulmonary fibrosis and emphysema (CPFE), asthma, and respiratory infections (eg, coronavirus infection (COVID-19)). These lung diseases are related to each other, and various symptoms appear mixed, so it is known that it is necessary to be careful in selecting a therapeutic agent. The main pathogenesis of lung disease includes lung injury and inflammatory response, and fibrosis.

Specifically, when an inflammatory reaction occurs in the lungs due to viruses, microorganisms, harmful substances, etc., inflammatory cytokines (eg, IL-1, IL-6, TNF-α) secretion by alveolar macrophages, attraction of neutrophils, It is known that the elasticity and structure of the lung tissue are damaged due to the secretion of protease (eg, elastase) that breaks down the fibers constituting the lung tissue and leads to fibrosis of the lungs. Inflammation and fibrosis of the lungs precede the progression of many lung diseases, so they can be said to be fundamental mechanisms in the prevention and treatment of lung diseases.

Pulmonary fibrosis is a representative example of lung disease, and fibrosis is a disease in which excessive fibrous connective tissue is formed in an organ or tissue. Fibrosis refers to a state in which normal control is impossible during the wound healing process after tissue is damaged by various stresses (infection, chemical stimulation, radiation, etc.) in the human body. Fibrosis occurs in various organs such as the lungs, heart, and liver, and is one of the fields with high unmet demand as no fundamental treatment has yet been developed.

Idiopathic pulmonary fibrosis (IPF), one of pulmonary fibrosis of unknown cause, is a chronic disease of fibrotic interstitial pneumonia of unknown etiology, in which fibrosis progresses due to continuous damage to alveolar epithelial cells. Treatment methods according to the administration of pirfenidone and nintedanib have been studied, but side effects such as decreased appetite, weakness, digestive side effects, hepatotoxicity potential, and photosensitivity rash are known.

In addition, chronic obstructive pulmonary disease (COPD), another representative example of lung disease, is a disease in which airway obstruction occurs gradually as the airway narrows due to an abnormal inflammatory response of the lungs by tobacco, air pollution, or toxic inhalation substances. It is broadly divided into chronic bronchitis and emphysema. In particular, smoking is known to be the main cause of chronic obstructive pulmonary disease. Smoking acts as a strong irritant in the lung tissue, increasing the production of various pro-inflammatory factors, growth factors, oxidizing substances and chemotactic factors, and activating the inflammatory signaling system, leading to migration of many inflammatory cells including neutrophils and macrophages. It promotes lung inflammation further aggravates lung inflammation, which in turn causes abnormal changes in lung tissue, such as airway wall thickening, lung fibrosis, and lowers lung function. Accordingly, improvement of lung inflammation is understood as a treatment method for the prevention and treatment of chronic obstructive pulmonary disease.

Another representative example of lung disease is lung damage caused by a new coronavirus (2019-nCoV or SARS-CoV-2) infection and acute respiratory disease (COVID-19). The lung is known as the main vulnerable organ because the novel coronavirus, transmitted through the respiratory tract, penetrates into the cell through ACE2 and TMPRSS2, which are mainly expressed in type II alveolar epithelial cells. The main symptoms include fever and cough, and healthy adults are more likely to recover over time. However, it is known that when the virus infection in the lungs is severe due to low initial immunity, it causes a serious inflammatory response according to lung damage and promotes fibrosis, which can be accompanied by symptoms such as acute respiratory distress syndrome (ARDS) and sepsis. . Currently, drugs that suppress infection and proliferation of viruses or control lung inflammation are mainly used for COVID-19 treatment. Accordingly, improvement of lung inflammation and fibrosis is understood as a treatment method for the prevention and treatment of COVID-19.

As such, since lung inflammation and fibrosis are major causes of the onset and development of lung diseases, improvement of lung inflammation and fibrosis has been studied as a therapeutic mechanism for various lung diseases.

On the other hand, GLP-1 (glucagon-like peptide-1) and GIP (glucose-dependent insuliontropic polypeptide) are representative gastrointestinal hormones and neurohormones, which are involved in the regulation of blood sugar components according to food intake. Glucagon is a peptide hormone secreted by the pancreas and is involved in blood sugar concentration control together with the two substances described above. The development of therapeutic agents using drugs that can each or simultaneously act on the GLP-1 receptor, the GIP receptor, and the glucagon receptor has been made (US 10,370,426, US 10,400,020).

Although various studies on lung diseases have been conducted so far, the development of a practical and effective therapeutic agent is still insufficient, and there is a need for continuous development of therapeutic agents.

One object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of pulmonary disease, comprising a peptide having activity against glucagon receptors, GLP-1 receptors and GIP receptors, or a long-acting conjugate of these peptides.

Another object of the present invention is to provide a method for preventing or treating lung disease, comprising administering to an individual in need thereof a composition comprising the peptide or a long-acting conjugate of the peptide.

Another object of the present invention is to provide the use of a composition comprising the peptide or a long-acting conjugate of the peptide in the manufacture of a medicament for the prevention or treatment of lung disease.

One aspect embodying the present invention is a glucagon receptor, GLP-1 (Glucagon-like peptide-1) receptor, and GIP (Glucose-dependent insulinotropic polypeptide) receptors for the prevention or treatment of lung diseases comprising a peptide having activity. a pharmaceutical composition for

In one embodiment, the pharmaceutical composition for the prevention or treatment of the lung disease comprises a pharmaceutically effective amount of a pharmaceutically acceptable excipient and a peptide comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 102. characterized.

The pharmaceutical composition according to any one of the preceding embodiments, wherein the peptide is in the form of a long-acting conjugate, and the long-acting conjugate is characterized in that it is represented by the following formula (1):

[Formula 1]

X - L - F

With the proviso that X is a peptide of any one of SEQ ID NOs: 1 to 102;

L is a linker containing an ethylene glycol repeating unit,

F is an immunoglobulin Fc region,

- represents a covalent linkage between X and L and between L and F.

The composition according to any one of the preceding embodiments, wherein the peptide is amidated at its C-terminus.

The composition according to any one of the preceding embodiments, wherein the peptide is characterized in that its C-terminus is amidated or has a free carboxyl group (-COOH).

The composition according to any one of the preceding embodiments, wherein the peptide is selected from the group consisting of SEQ ID NOs: 21, 22, 42, 43, 50, 64, 66, 67, 70, 71, 76, 77, 96, 97 and 100 It is characterized in that it contains amino acids.

The composition according to any one of the preceding embodiments, wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 21, 22, 42, 43, 50, 77 and 96.

The composition according to any one of the preceding embodiments, wherein the peptide sequence is characterized in that amino acids 16 and 20 from the N-terminus form a ring with each other.

The composition according to any one of the preceding embodiments, wherein the formula weight of the ethylene glycol repeating unit moiety in L is in the range of 1 to 100 kDa.

The composition according to any one of the preceding embodiments, wherein F is an IgG Fc region.

The composition according to any one of the preceding embodiments, wherein the lung disease is interstitial lung disease (ILD), progressive fibrosing interstitial lung disease (PF-ILD), idiopathic interstitial pneumonia ( idiopathic interstitial pneumonias (IIP), non-specific interstitial pneumonia (NSIP), pulmonary fibrosis, fibrosing interstitial lung diseases (FILD), idiopathic pulmonary fibrosis, IPF), alveolitis, pneumonia, emphysema, bronchitis, chronic obstructive pulmonary disease, combined pulmonary fibrosis and emphysema (CPFE), It is characterized in that it is asthma, or a respiratory infection.

The composition according to any one of the preceding embodiments, wherein the respiratory infectious disease is a respiratory viral, bacterial, mycoplasma, or fungal infection.

The composition according to any one of the preceding embodiments, wherein the respiratory virus is adenovirus, vaccinia virus, herpes simplex virus, parainfluenza virus, rhinovirus ), varicella Zoster Virus, measle virus, respiratorysyncytial virus, Dengue virus, HIV (human immunodeficiency virus), influenza virus, coronavirus, severe It is characterized in that it is any one selected from the group consisting of severe acute respiratory syndrome associated virus (SARS-associated virus), and middle east respiratory syndrome coronavirus (MERS-CoV).

The composition according to any one of the preceding embodiments, wherein the coronavirus is SARS-CoV-2.

The composition according to any one of the preceding embodiments, wherein, upon administration, the pharmaceutical composition (i) inhibits macrophage activity, and/or (ii) IL-1β, IL-6, IL-12, or TNF- It is characterized by reducing the expression of α.

A composition according to any one of the preceding embodiments, wherein the pharmaceutical composition has at least one of the following properties upon administration:

(i) inhibition of myofibroblast differentiation;

(ii) decreased expression of α-SMA, collagen1α1, or fibronectin; or

(iii) inhibition of epithelial mesenchymal transition (EMT) of alveolar epithelial cells; and

(iv) decreased expression of collagen1α1, or collagen1α3

The composition according to any one of the preceding embodiments, wherein the pharmaceutical composition is characterized in that mucolytic agents or pharmaceutically acceptable salts thereof are additionally administered.

The composition according to any one of the preceding embodiments, wherein the mucolytic agent is ambroxol, N-acetylcysteine, N-acetylin, carbocysteine, sea bream. domiodol, fudosteine, bromhexine, erdosteine, letostine, lysozyme, mesna, sobrerol, stefronin (stepronin), thiopronin, tyloxapol, carbocisteine, dornase alfa, eprazinone, letosteine, neltenexine, and at least one selected from the group consisting of mecysteine.

The composition according to any one of the preceding embodiments, wherein the peptide and the mucolytic agent or a pharmaceutically acceptable salt thereof are administered simultaneously, sequentially, or in reverse order.

The composition according to any one of the preceding embodiments, wherein the lung disease may be characterized in that the lung inflammation and fibrosis caused by coronavirus infection-19 (COVID-19).

The composition according to any one of the preceding embodiments, wherein the region F is a dimer consisting of two polypeptide chains, and one end of L is linked to only one of the two polypeptide chains.

Another aspect embodying the present invention is a method for preventing or treating lung disease, comprising administering the peptide or a composition comprising the same to an individual in need thereof.

Another aspect embodying the present invention is the use of the peptide or a composition comprising the same in the manufacture of a medicament for the prevention or treatment of lung disease.

Another aspect embodying the present invention is the use of the peptide or a composition comprising the same for the prevention or treatment of lung disease.

The triple activator or a long-acting conjugate thereof according to the present invention has activity against a glucagon receptor, a GLP-1 (Glucagon-like peptide-1) receptor, and a GIP (glucose-dependent insulinotropic polypeptide) receptor, thereby preventing lung disease Or it may exhibit a therapeutic effect.

1 is a view confirming the change in the level of inflammatory cytokine expression in the lung tissue according to the triple activator long-acting conjugate treatment in vivo.
2 is It is a diagram confirming the effect of improving emphysema in vivo by treatment with the triple active agent long-acting conjugate.
Figure 3 is a view confirming the change in the expression level of myofibroblast differentiation markers (α-SMA, collagen1α1, fibronectin) in lung fibroblasts (lung fibroblast, MRC5 cell) according to the treatment of the triple activator long-acting conjugate.
Figure 4 is a diagram confirming the change in the expression level of epithelial mesenchymal transition (EMT) markers (collagen1α1, collagen1α3) in alveolar epithelial cells (lung alveolar epithelial cells, A549 cells) according to the treatment of the triple activator long-acting conjugate .
5 is a diagram confirming in vivo the effect of improving the fibrosis of BLM mouse lung tissue by treatment with the triple activator long-acting conjugate.
6 is a view confirming the change in the survival rate of BLM mice according to the triple activator long-acting conjugate treatment.

Hereinafter, the present invention will be described in more detail.

On the other hand, each description and embodiment disclosed herein may be applied to each other description and embodiment. That is, all combinations of the various elements disclosed herein are within the scope of the present invention. In addition, it cannot be said that the scope of the present invention is limited by the specific descriptions described below.

Throughout this specification, common one-letter and three-letter codes for naturally occurring amino acids are used as well as Aib (2-aminoisobutyric acid, 2-aminoisobutyric acid), Sar (N-methylglycine), alpha-methyl A commonly accepted three-letter code is used for other amino acids, such as α-methyl-glutamic acid. Amino acids referred to by abbreviation herein are also described according to the IUPAC-IUB nomenclature.

alanine Ala, A arginine Arg, R

asparagine Asn, N Aspartic Acid Asp, D

cysteine Cys, C glutamic acid Glu, E

glutamine Gln, Q glycine Gly, G

histidine His, H isoleucine Ile, I

leucine Leu, L. Lee Sin Lys, K.

methionine Met, M. phenylalanine Phe, F.

proline Pro, P serine Ser, S

threonine Thr, T tryptophan Trp, W

tyrosine Tyr, Y valine Val, V

One aspect for implementing the present invention is a glucagon receptor, GLP-1 (Glucagon-like peptide-1) receptor, and GIP (Glucose-dependent insulinotropic polypeptide) having activity on the receptor, including a peptide, of lung disease It is a pharmaceutical composition for prevention or treatment.

In one embodiment, the peptide may include the amino acid sequence of any one of SEQ ID NOs: 1 to 102.

In another embodiment, the pharmaceutical composition for the prevention or treatment of lung disease is a pharmaceutical comprising a pharmaceutically acceptable excipient and a peptide comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 102 in a pharmaceutically effective amount It may be an enemy composition.

The term “peptide having activity against glucagon receptor, GLP-1 receptor, and GIP receptor” may be used interchangeably as the term “triple activator” in the present invention.

Such peptides include various substances with significant levels of activity on glucagon, GLP-1, and GIP receptors, such as various peptides.

Although not particularly limited thereto, the triple activator having a significant level of activity on the glucagon, GLP-1, and GIP receptors includes one or more receptors of glucagon, GLP-1, and GIP receptors, specifically two or more More specifically, the in vitro activity for all three receptors is about 0.001% or more, about 0.01% or more, compared to the native ligand (natural glucagon, native GLP-1, and native GIP) of the corresponding receptor, about 0.1% or more, about 1% or more, about 2% or more, about 3% or more, about 4% or more, about 5% or more, about 6% or more, about 7% or more, about 8% or more, about 9% or more, About 10% or more, about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, about 100% or more However, the significantly increased range is included without limitation.

Here, the activity of the receptors, the in vitro activity against wild-type compared to the receptor at least 0.1%, at least 1%, 2% or more, at least 3%, more than 4%, 5%, 6%, 7% or more, 8% or more, 9% or more, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, about 200 % or more can be exemplified. However, it is not limited thereto.

In the present invention, the term "about" includes all ranges including ±0.5, ±0.4, ±0.3, ±0.2, ±0.1, etc., and includes all values in a range equal to or similar to the value following the term about, but not limited

For a method of measuring the in vitro activity of such a triple activator, reference may be made to Experimental Example 1 of the present specification, but is not particularly limited thereto.

On the other hand, the peptide possesses one or more, two or more, specifically three activities of the following i) to iii), specifically, it is characterized in that it possesses a significant activity:

i) activation of the GLP-1 receptor; ii) activation of the glucagon receptor; and iii) activation of the GIP receptor.

Here, activating the receptor means that the in vitro activity of the receptor compared to the native type is about 0.1% or more, about 1% or more, about 2% or more, about 3% or more, about 4% or more, about 5% or more, about 6 % or more, about 7% or more, about 8% or more, about 9% or more, about 10% or more, about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70 % or more, about 80% or more, about 90% or more, about 100% or more may be exemplified. However, it is not limited thereto.

In addition, the peptide may have an increased half-life in the body compared to any one of native GLP-1, native glucagon, and native GIP, but is not particularly limited thereto.

Although not particularly limited thereto, the peptide may be non-naturally occurring.

The peptide may be an analog of native glucagon, but is not particularly limited thereto. Specifically, the native glucagon analog includes peptides having one or more differences in amino acid sequence compared to native glucagon, peptides modified through modification of the native glucagon sequence, and mimics of native glucagon.

Meanwhile, although not particularly limited thereto, native glucagon may have the following amino acid sequence:

His-Ser-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Ser-Arg-Arg-Ala-Gln-Asp-Phe-Val-Gln-Trp- Leu-Met-Asn-Thr (SEQ ID NO: 118)

Specifically, the peptide has at least one amino acid in the native glucagon sequence in which a modification selected from the group consisting of substitution, addition, deletion, modification, and combinations thereof has occurred. It may be an analog of glucagon, but is not particularly limited thereto.

In addition, the substitution of amino acids includes both substitutions with amino acids and substitutions with non-natural compounds.

Additionally, additions may be made at the N-terminus and/or C-terminus of the peptide. On the other hand, the length of the amino acid to be added is not particularly limited thereto, and 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more amino acids may be added. and can broadly include the addition of a polypeptide, but is not particularly limited thereto.

More specifically, the peptide is 1, 2, 3, 7, 10, 12, 13, 14, 15, 16, 17, 18, 19 in the native glucagon amino acid sequence. times, 20 times, 21 times, 23 times, 24 times, 27 times, 28 times and 29 times, 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 More than, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 or more, or 20 amino acids may be substituted with other amino acids, and Independently or additionally, 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more amino acids may be added to the C-terminus thereof, It is not particularly limited thereto.

More specifically, the peptide is 1, 2, 3, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20 in the native glucagon amino acid sequence. 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 More than, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 amino acids may be substituted with other amino acids, and also independently or additionally its C- 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, or 11 or more amino acids may be added to the terminal, but is not particularly limited thereto .

More specifically, the peptide is 1, 2, 3, 10, 13, 14, 15, 16, 17, 18, 19, 20, 21 in the native glucagon amino acid sequence. 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 More than, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 amino acids may be substituted with other amino acids, and independently or additionally 1 or more, 2 or more, 3 or more, 4 or more at the C-terminus thereof More than, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, or 11 or more amino acids may be added, but is not particularly limited thereto.

More specifically, the peptide is 1, 2, 13, 16, 17, 18, 19, 20, 21, 23, 24, 27, 28 in the native glucagon amino acid sequence. 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, or 14 amino acids may be substituted with other amino acids, and independently or additionally 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, at the C-terminus thereof, 10 or more, 11 or more amino acids may be added, but is not particularly limited thereto.

The introduced amino acid is from the group consisting of tyrosine, alpha-methyl-glutamic acid, Aib, methionine, glutamic acid, histidine, lysine, leucine, isoleucine, glutamine, valine, glycine, alanine, cysteine, serine, alanine, aspartic acid, and arginine. may be selected, but is not particularly limited thereto.

For example, the added amino acid sequence may be one or more amino acid sequences derived from a native GLP-1, native GIP, or native exendin-4 amino acid sequence.

Such a peptide may include an intramolecular bridge (eg, a covalent bridge or a non-covalent bridge), and specifically may be in a form containing a ring, for example, between amino acids 16 and 20 of the peptide. It may be in a form in which a ring is formed, but is not particularly limited thereto.

Non-limiting examples of the ring may include a lactam bridge (or lactam ring).

In addition, the peptide includes all those modified to include an amino acid capable of forming a ring at a desired position to include a ring.

For example, the pair of amino acids 16 and 20 of the peptide may be substituted with glutamic acid or lysine, each capable of forming a ring, but is not limited thereto.

Such a ring may be formed between amino acid side chains in the peptide, for example, a lactam ring may be formed between a lysine side chain and a glutamic acid side chain, but is not particularly limited thereto.

As an example of a peptide prepared by a combination of these methods, the amino acid sequence differs from native glucagon by one or more, and the alpha-carbon of the amino acid residue at the N-terminus has been removed, glucagon receptor, GLP-1 receptor, and activity against GIP receptor There are peptides having a . However, the present invention is not limited thereto, and a combination of various methods for analog production can be used to prepare the peptides applied to the present invention.

In addition, although not particularly limited thereto, in the peptide of the present invention, some amino acids may be substituted with other amino acids or non-natural compounds in order to avoid the recognition action of an activator degrading enzyme in order to increase the half-life in the body.

Specifically, the peptide may be a peptide having an increased half-life in the body by avoiding the recognition action of the degrading enzyme through substitution of the second amino acid sequence among the amino acid sequence of the peptide, but amino acid substitution or alteration to avoid the recognition action of the degrading enzyme in the body included without limitation.

In addition, such modifications for the preparation of peptides include modifications with L- or D-form amino acids, and/or non-naturally occurring amino acids; and/or modifying the native sequence by modifying, for example, modification of side chain functional groups, intramolecular covalent bonds, such as inter-side chain ring formation, methylation, acylation, ubiquitination, phosphorylation, aminohexylation, biotinylation, etc. includes all that

In addition, it includes all those in which one or more amino acids are added to the amino and/or carboxy terminus of native glucagon.

The amino acids to be substituted or added may be atypical or non-naturally occurring amino acids as well as the 20 amino acids commonly found in human proteins. Commercial sources of atypical amino acids include Sigma-Aldrich, ChemPep and Genzyme Pharmaceuticals. Peptides containing these amino acids and canonical peptide sequences can be synthesized and purchased from commercial peptide synthesis companies, for example, American peptide company or Bachem in the United States, or Anygen in Korea.

Amino acid derivatives can also be obtained in the same way, and to name just a few examples, 4-imidazoacetic acid and the like can be used.

In addition, the peptide according to the present invention is protected from proteolytic enzymes in vivo and its N-terminus and/or C-terminus is chemically modified or protected by an organic group, or amino acids are added to the peptide terminus to increase stability. It may be added and modified form.

In particular, in the case of a chemically synthesized peptide, since the N- and C-terminals are charged, the N-terminus is acetylated and/or the C-terminus is amidated to remove these charges. However, it is not particularly limited thereto.

In addition, the peptide according to the present invention includes all forms of the peptide itself, a salt thereof (eg, a pharmaceutically acceptable salt of the peptide), or a solvate thereof. In addition, the peptide may be in any pharmaceutically acceptable form.

The type of the salt is not particularly limited. However, it is preferably in a form that is safe and effective for an individual, such as a mammal, but is not particularly limited thereto.

The term, "pharmaceutically acceptable" means a material that can be effectively used for a desired purpose without causing excessive toxicity, irritation, or allergic reaction within the scope of medical judgment.

As used herein, the term "pharmaceutically acceptable salt" includes salts derived from pharmaceutically acceptable inorganic acids, organic acids, or bases. Examples of suitable acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid , benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include alkali metals such as sodium and potassium, alkaline earth metals such as magnesium, and ammonium.

In addition, as used herein, the term "solvate" refers to a compound in which the peptide or a salt thereof according to the present invention forms a complex with a solvent molecule.

In one embodiment, the peptide may include an amino acid sequence represented by the following general formula (1).

Xaa1-Xaa2-Xaa3-Gly-Thr-Phe-Xaa7-Ser-Asp-Xaa10-Ser-Xaa12-Xaa13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-Xaa19-Xaa20-Xaa21-Phe-Xaa23-Xaa24-Trp23 Leu-Xaa27-Xaa28-Xaa29-Xaa30-R1 (General Formula 1, SEQ ID NO: 103)

In the above general formula 1,

Xaa1 is histidine, 4-imidazoacetyl, or tyrosine;

Xaa2 is glycine, alpha-methyl-glutamic acid, or Aib;

Xaa3 is glutamic acid or glutamine,

Xaa7 is threonine or isoleucine,

Xaa10 is leucine, tyrosine, lysine, cysteine, or valine;

Xaa12 is lysine, serine, or isoleucine,

Xaa13 is glutamine, tyrosine, alanine, or cysteine,

Xaa14 is leucine, methionine, or tyrosine,

Xaa15 is cysteine, aspartic acid, glutamic acid, or leucine,

Xaa16 is glycine, glutamic acid, or serine,

Xaa17 is glutamine, arginine, isoleucine, glutamic acid, cysteine, or lysine;

Xaa18 is alanine, glutamine, arginine, or histidine;

Xaa19 is alanine, glutamine, cysteine, or valine,

Xaa20 is lysine, glutamine, or arginine;

Xaa21 is glutamic acid, glutamine, leucine, cysteine, or aspartic acid;

Xaa23 is isoleucine or valine,

Xaa24 is alanine, glutamine, cysteine, asparagine, aspartic acid, or glutamic acid,

Xaa27 is valine, leucine, or lysine;

Xaa28 is cysteine, lysine, alanine, asparagine, or aspartic acid;

Xaa29 is cysteine, glycine, glutamine, threonine, glutamic acid, or histidine;

Xaa30 is cysteine, glycine, lysine, or histidine, or is absent;

R1 is cysteine, GKKNDWKHNIT (SEQ ID NO: 106), m-SSGAPPPS-n (SEQ ID NO: 107), or m-SSGQPPPS-n (SEQ ID NO: 108), or is absent;

here,

m is -Cys-, -Pro-, or -Gly-Pro-;

n is -Cys-, -Gly-, -Ser-, or -His-Gly-, or absent.

Examples of the triple activator include those comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 102, SEQ ID NOs: 1 to 11, those comprising an amino acid sequence selected from the group consisting of 13 to 102, SEQ ID NOs: 1 to 11, and may be (essentially) composed of an amino acid sequence selected from the group consisting of 13 to 102, but is not limited thereto.

In another embodiment, the triple activator has an amino acid sequence selected from the group consisting of SEQ ID NOs: 21, 22, 42, 43, 50, 64, 66, 67, 70, 71, 76, 77, 96, 97 and 100 It may be (essentially) composed of, but is not limited thereto.

In another embodiment, the triple activator may be (essentially) composed of an amino acid sequence selected from the group consisting of SEQ ID NOs: 21, 22, 42, 43, 50, 66, 67, 77, 96, 97 and 100 However, the present invention is not limited thereto.

In another embodiment, the triple activator may be (essentially) composed of an amino acid sequence selected from the group consisting of SEQ ID NOs: 21, 22, 42, 43, 50, 77 and 96, but is not limited thereto.

In addition, even if it is described herein as a peptide 'consisting of a specific SEQ ID NO: If it has the same or corresponding activity as the peptide consisting of the amino acid sequence of the corresponding SEQ ID NO: Addition of meaningless sequences before and after the amino acid sequence of the corresponding SEQ ID NO: Naturally occurring mutations or latent mutations thereof are not excluded, and it is apparent that such sequence additions or mutations fall within the scope of the present application.

The above content may be applied to other embodiments or other aspects of the present invention, but is not limited thereto.

Specifically, in Formula 1, Xaa14 may be leucine or methionine, and Xaa15 may be cysteine, aspartic acid, or leucine.

Examples of such a peptide include, but are not particularly limited to, a peptide comprising or (essentially) consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 11, 14 to 17, and 21 to 102.

Such a peptide may significantly activate one or more of a glucagon receptor, a GLP-1 receptor, and a GIP receptor, but is not particularly limited thereto. Specifically, it may significantly activate GLP-1, or further significantly activate glucagon receptor and/or GIP receptor, but is not particularly limited thereto.

More specifically,

In the above general formula 1,

Xaa2 is glycine, alpha-methyl-glutamic acid, or Aib;

Xaa7 is threonine,

Xaa10 is tyrosine, cysteine, or valine,

Xaa12 is lysine or isoleucine,

Xaa13 is tyrosine, alanine, glutamine, or cysteine,

Xaa14 is leucine, cysteine, or methionine,

Xaa15 is cysteine, leucine, glutamic acid, or aspartic acid,

Xaa17 is glutamine, arginine, isoleucine, cysteine, glutamic acid, or lysine,

Xaa18 is alanine, glutamine, arginine, or histidine;

Xaa19 is alanine, glutamine, valine, or cysteine,

Xaa20 is lysine, arginine, or glutamine;

Xaa21 is glutamic acid, glutamine, leucine, cysteine, or aspartic acid;

Xaa23 is isoleucine or valine,

Xaa24 is cysteine, alanine, glutamine, asparagine, glutamic acid, or aspartic acid;

Xaa27 may be a peptide, which is leucine or lysine, but is not particularly limited thereto.

More specifically,

In the above general formula 1,

Xaa2 is glycine, alpha-methyl-glutamic acid, or Aib;

Xaa7 is threonine,

Xaa10 is tyrosine, cysteine, or valine,

Xaa12 is lysine or isoleucine,

Xaa13 is tyrosine, alanine, or cysteine,

Xaa14 is leucine or methionine,

Xaa15 is cysteine or aspartic acid,

Xaa17 is glutamine, arginine, isoleucine, cysteine, or lysine,

Xaa18 is alanine, arginine, or histidine;

Xaa19 is alanine, glutamine, or cysteine,

Xaa20 is lysine or glutamine,

Xaa21 is glutamic acid, cysteine, or aspartic acid,

Xaa23 is valine,

Xaa24 is alanine, glutamine, cysteine, asparagine, or aspartic acid;

Xaa27 may be leucine or lysine, but is not particularly limited thereto.

More specifically,

In the above general formula 1,

Xaa2 is alpha-methyl-glutamic acid or Aib,

Xaa7 is threonine,

Xaa10 is tyrosine or cysteine,

Xaa12 is lysine or isoleucine,

Xaa13 is tyrosine, alanine, or cysteine,

Xaa14 is leucine or methionine,

Xaa15 is cysteine or aspartic acid,

Xaa16 is glutamic acid,

Xaa17 is arginine, isoleucine, cysteine, or lysine,

Xaa18 is alanine, arginine, or histidine;

Xaa19 is alanine, glutamine, or cysteine,

Xaa20 is lysine or glutamine,

Xaa21 is glutamic acid or aspartic acid,

Xaa23 is valine,

Xaa24 is glutamine, asparagine, or aspartic acid;

Xaa27 is leucine,

Xaa28 may be cysteine, alanine, asparagine, or aspartic acid.

Specifically,

In the above general formula 1,

Xaa1 is histidine or 4-imidazoacetyl,

Xaa2 is alpha-methyl-glutamic acid or Aib,

Xaa3 is glutamine,

Xaa7 is threonine,

Xaa10 is tyrosine,

Xaa12 is isoleucine,

Xaa13 is alanine or cysteine,

Xaa14 is methionine,

Xaa15 is aspartic acid,

Xaa16 is glutamic acid,

Xaa17 is isoleucine or lysine,

Xaa18 is alanine or histidine,

Xaa19 is glutamine or cysteine,

Xaa20 is lysine,

Xaa21 is aspartic acid,

Xaa23 is valine,

Xaa24 is asparagine,

Xaa27 is leucine,

Xaa28 is alanine or asparagine,

Xaa29 is glutamine or threonine,

Xaa30 may be cysteine or lysine or absent.

More specifically,

In the above general formula 1,

Xaa2 is glycine, alpha-methyl-glutamic acid, or Aib;

Xaa3 is glutamine,

Xaa7 is threonine,

Xaa10 is tyrosine, cysteine, or valine,

Xaa12 is lysine,

Xaa13 is tyrosine,

Xaa14 is leucine,

Xaa15 is aspartic acid,

Xaa16 is glycine, glutamic acid, or serine,

Xaa17 is glutamine, arginine, cysteine, or lysine,

Xaa18 is alanine, arginine, or histidine;

Xaa19 is alanine or glutamine,

Xaa20 is lysine or glutamine,

Xaa21 is glutamic acid, cysteine, or aspartic acid,

Xaa23 is valine,

Xaa24 is alanine, glutamine, or cysteine,

Xaa27 is leucine or lysine,

Xaa29 may be glycine, glutamine, threonine, or histidine, but is not particularly limited thereto.

These peptides have significant activation levels of GLP-1 receptors and glucagon receptors, and are higher than those of GIP receptors; The degree of activation of the GLP-1 receptor, the glucagon receptor and the GIP receptor is all significant; The degree of activation of the GLP-1 receptor and the GIP receptor is significant and may correspond to a case where the activation level of the glucagon receptor is higher than that of the glucagon receptor, but is not particularly limited thereto.

Examples of such a peptide include SEQ ID NOs: 8, 9, 21 to 37, 39, 42, 43, 49 to 61, 64 to 83, 85, 86, 88, 89, 91 to 93, selected from the group consisting of 95 to 102 and a peptide comprising or (essentially) consisting of an amino acid sequence, but is not particularly limited thereto.

In a specific embodiment, the peptide may include an amino acid sequence represented by the following general formula (2).

Xaa1-Xaa2-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Xaa10-Ser-Lys-Xaa13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-Xaa19-Xaa20-Xaa21-Phe-Xaa23-Xaa24-Trp Leu-Leu-Xaa28-Xaa29-Xaa30-Xaa31- Ser-Ser-Gly-Gln-Pro-Pro-Pro-Ser-Xaa40 (Formula 2, SEQ ID NO: 104)

In the above formula,

Xaa1 is 4-imidazoacetyl, histidine, or tyrosine;

Xaa2 is glycine, alpha-methyl-glutamic acid, or Aib;

Xaa10 is tyrosine or cysteine

Xaa13 is alanine, glutamine, tyrosine, or cysteine;

Xaa14 is leucine, methionine, or tyrosine;

Xaa15 is aspartic acid, glutamic acid, or leucine;

Xaa16 is glycine, glutamic acid, or serine;

Xaa17 is glutamine, arginine, isoleucine, glutamic acid, cysteine, or lysine;

Xaa18 is alanine, glutamine, arginine, or histidine;

Xaa19 is alanine, glutamine, cysteine, or valine;

Xaa20 is lysine, glutamine, or arginine;

Xaa21 is cysteine, glutamic acid, glutamine, leucine, or aspartic acid;

Xaa23 is isoleucine or valine;

Xaa24 is cysteine, alanine, glutamine, asparagine, or glutamic acid;

Xaa28 is lysine, cysteine, asparagine, or aspartic acid;

Xaa29 is glycine, glutamine, cysteine, or histidine;

Xaa30 is cysteine, glycine, lysine, or histidine;

Xaa31 is proline or cysteine;

Xaa40 is cysteine or absent.

More specifically, in the general formula 2,

Xaa13 is alanine, tyrosine, or cysteine;

Xaa15 is aspartic acid or glutamic acid,

Xaa17 is glutamine, arginine, cysteine, or lysine;

Xaa18 is alanine, arginine, or histidine;

Xaa21 is cysteine, glutamic acid, glutamine, or aspartic acid;

Xaa23 is isoleucine or valine;

Xaa24 is cysteine, glutamine, or asparagine;

Xaa28 is cysteine, asparagine, or aspartic acid;

Xaa29 is glutamine, cysteine, or histidine;

Xaa30 may be cysteine, lysine, or histidine.

Examples of such a peptide include an amino acid sequence selected from the group consisting of SEQ ID NOs: 21, 22, 42, 43, 50, 64 to 77, and 95 to 102, more specifically SEQ ID NOs: 21, 22, 42, 43, 50, 64 to 77, and a peptide comprising or (essentially) consisting of an amino acid sequence selected from the group consisting of 96 to 102, but is not particularly limited thereto.

In a specific embodiment, the peptide may include an amino acid sequence of the following general formula (3).

Xaa1-Xaa2-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Xaa13-Leu-Asp-Glu-Xaa17-Xaa18-Xaa19-Lys-Xaa21-Phe-Val-Xaa24-Trp- Leu-Leu-Xaa28-Xaa29-Xaa30-Xaa31-Ser-Ser-Gly-Gln-Pro-Pro-Pro-Ser-Xaa40 (Formula 3, SEQ ID NO: 105);

In the above general formula 3,

Xaa1 is histidine or tyrosine;

Xaa2 is alpha-methyl-glutamic acid or Aib;

Xaa13 is alanine, tyrosine or cysteine;

Xaa17 is arginine, cysteine, or lysine;

Xaa18 is alanine or arginine;

Xaa19 is alanine or cysteine;

Xaa21 is glutamic acid or aspartic acid;

Xaa24 is glutamine or asparagine,

Xaa28 is cysteine or aspartic acid;

Xaa29 is cysteine, histidine, or glutamine;

Xaa30 is cysteine or histidine;

Xaa31 is proline or cysteine;

Xaa40 may be cysteine or absent.

Examples of such a peptide include a peptide comprising or (essentially) consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 21, 22, 42, 43, 50, 64 to 71, 75 to 77, and 96 to 102. However, it is not particularly limited thereto.

In addition, in Formula 1, R1 is cysteine, GKKNDWKHNIT (SEQ ID NO: 106), CSSGQPPPS (SEQ ID NO: 109), GPSSGAPPPS (SEQ ID NO: 110), GPSSGAPPPSC (SEQ ID NO: 111), PSSGAPPPS (SEQ ID NO: 112), PSSGAPPPSG (SEQ ID NO: SEQ ID NO: 110) 113), PSSGAPPPSHG (SEQ ID NO: 114), PSSGAPPPSS (SEQ ID NO: 115), PSSGQPPPS (SEQ ID NO: 116), or PSSGQPPPSC (SEQ ID NO: 117), or absent, but is not particularly limited thereto.

In addition, according to the length of the peptide of the present invention, it can be synthesized by a method well known in the art, for example, an automatic peptide synthesizer, or it can be produced by a genetic engineering technique.

Specifically, the peptides of the present invention can be prepared by standard synthetic methods, recombinant expression systems, or any other method in the art. Thus, the peptides according to the invention can be synthesized in a number of ways, including, for example, those comprising:

(a) synthesizing peptides stepwise or by fragment assembly by means of solid or liquid phase methods, and isolating and purifying the final peptide product; or

(b) expressing a nucleic acid construct encoding the peptide in a host cell and recovering the expression product from the host cell culture; or

(c) performing cell-free in vitro expression of a nucleic acid construct encoding the peptide and recovering the expression product; or

A method for obtaining a fragment of a peptide by any combination of (a), (b) and (c), and then ligating the fragments to obtain a peptide, and recovering the peptide.

In addition, the peptide having activity on the glucagon receptor, the GLP-1 receptor, and the GIP receptor is biocompatible to increase the in vivo half-life of the peptide having activity on the glucagon receptor, the GLP-1 receptor, and the GIP receptor. The substance may be in the form of a bound, long-acting binder. In the present specification, the biocompatible material may be mixed with a carrier.

In the present invention, the conjugate of the peptide may exhibit increased potency and durability compared to the peptide to which the carrier is not bound, and in the present invention, such a conjugate is referred to as a "persistent conjugate".

Meanwhile, the combination may be non-naturally occurring.

In one embodiment of the present invention, the long-acting conjugate may be one represented by the following formula (1), but is not limited thereto:

[Formula 1]

X - L - F

with the proviso that X is a peptide comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 102;

L is a linker containing ethylene glycol repeating units,

F is an immunoglobulin Fc fragment or derivative thereof,

- represents a covalent linkage between X and L and between L and F.

In the conjugate, F is X, that is, a peptide having activity on the glucagon receptor, the GLP-1 receptor, and the GIP receptor, specifically, the half-life of a peptide comprising any one of the amino acid sequences of SEQ ID NOs: 1 to 102. As a material that can be used, it corresponds to one component of the moiety constituting the binder of the present invention.

The F may be bonded to each other through a covalent chemical bond or a non-covalent chemical bond with X, and F and X may be bonded to each other through L through a covalent chemical bond, a non-covalent chemical bond, or a combination thereof.

Specifically, L may be a non-peptidyl linker, for example, a linker containing an ethylene glycol repeating unit.

In the present invention, "non-peptidyl linker" includes a biocompatible polymer in which two or more repeating units are bonded. The repeating units are linked to each other through any covalent bond other than a peptide bond. The non-peptidyl linker may be one component constituting a moiety of the conjugate of the present invention, and corresponds to L in Formula 1 above.

The non-peptidyl linker that can be used in the present invention may be used without limitation as long as it is a polymer resistant to in vivo protease. In the present invention, the non-peptidyl linker may be used in combination with a non-peptidyl polymer.

Although not particularly limited thereto, the non-peptidyl linker may be a linker containing an ethylene glycol repeating unit, for example, polyethylene glycol, and also derivatives thereof known in the art and easily at the level of skill in the art. Derivatives that can be prepared are also included in the scope of the present invention.

The repeating unit of the non-peptidyl linker may be an ethylene glycol repeating unit, and specifically, the non-peptidyl linker may include an ethylene glycol repeating unit and a functional group used in the preparation of the conjugate at the terminal thereof. The long-acting conjugate according to the present invention may be in a form in which X and F are linked through the functional group, but is not limited thereto. In the present invention, the non-peptidyl linker may include two, or three or more functional groups, and each functional group may be the same or different from each other, but is not limited thereto.

Specifically, the linker may be polyethylene glycol (PEG) represented by the following formula (2), but is not limited thereto:

[Formula 2]

Here, n=10 to 2400, n=10 to 480, or n=50 to 250, but is not limited thereto.

The PEG moiety in the long-acting conjugate may include _{not only the -(CH 2} CH ₂ O) _n -structure, but also an oxygen atom intervening between the linking element and the -(CH ₂ CH ₂ O) _{n -, but this} It is not limited.

In addition, in one specific embodiment, the conjugate may have a structure in which the peptide (X) comprising the amino acid sequence of Formula 1 and the immunoglobulin Fc region (F) are covalently linked through a linker containing an ethylene glycol repeating unit. However, the present invention is not limited thereto.

The polyethylene glycol is a term that encompasses all forms of ethylene glycol homopolymer, PEG copolymer, or monomethyl-substituted PEG polymer (mPEG), but is not particularly limited thereto.

The non-peptidyl linker that can be used in the present invention may be used without limitation as long as it is a polymer including an ethylene glycol repeating unit that is resistant to proteolytic enzymes in vivo. The molecular weight of the non-peptidyl polymer is, but is not limited to, greater than 0 in the range of about 100 kDa, in the range of about 1 to about 100 kDa, specifically in the range of about 1 to about 20 kDa, or in the range of about 1 to about 10 kDa. In addition, as the non-peptidyl linker of the present invention coupled to the polypeptide corresponding to F, not only one type of polymer but also a combination of different types of polymers may be used.

In one specific embodiment, both ends of the non-peptidyl linker may be bound to an amine or thiol group of F, such as an immunoglobulin Fc region, and an amine or thiol group of X, respectively.

Specifically, the non-peptidyl polymer has a reactive group capable of binding to F (eg, immunoglobulin Fc region) and X at both ends, specifically, the N-terminus of X, or F (eg, immunoglobulin Fc region). Or it may include a reactive group capable of bonding to an amine group located on lysine, or a thiol group of cysteine, but is not limited thereto.

In addition, the reactive group of the non-peptidyl polymer capable of binding to F, such as an immunoglobulin Fc region and X, may be selected from the group consisting of an aldehyde group, a maleimide group and a succinimide derivative, but is not limited thereto.

In the above, the aldehyde group may be exemplified by a propion aldehyde group or a butyl aldehyde group, but is not limited thereto.

In the above, as the succinimide derivative, succinimidyl valerate, succinimidyl methylbutanoate, succinimidyl methylpropionate, succinimidyl butanoate, succinimidyl propionate, N-hydroxysuccini Mead, hydroxy succinimidyl, succinimidyl carboxymethyl or succinimidyl carbonate may be used, but not limited thereto.

The non-peptidyl linker may be connected to X and F through such a reactive group, but is not particularly limited thereto.

In addition, the final product resulting from reductive amination by aldehyde bonds is much more stable than those linked by amide bonds. The aldehyde reactive group selectively reacts with the N-terminus at a low pH, and can form a covalent bond with a lysine residue at a high pH, for example, pH 9.0.

In addition, the reactive groups at both ends of the non-peptidyl linker may be the same or different from each other, for example, a maleimide group at one end and an aldehyde group, a propionaldehyde group, or a butyl aldehyde group at the other end. . However, if F, specifically, an immunoglobulin Fc region and X can be bound to each end of the non-peptide linker, it is not particularly limited thereto.

For example, one end of the non-peptidyl linker may include a maleimide group as a reactive group, and an aldehyde group, a propionaldehyde group, or a butyl aldehyde group at the other end of the non-peptidyl linker.

When polyethylene glycol having a hydroxyl reactive group at both ends is used as a non-peptidyl polymer, the hydroxyl group can be activated into the various reactive groups by a known chemical reaction, or a commercially available polyethylene glycol having a modified reactive group is used. The long-acting protein conjugate of the invention can be prepared.

In one specific embodiment, the non-peptidyl polymer may be linked to a cysteine residue of X, more specifically, a -SH group of cysteine, but is not limited thereto.

For example, in the peptide corresponding to X, cysteine residue 10, cysteine 13, cysteine 15, cysteine 17, cysteine 19, cysteine 21, cysteine 24, cysteine 28, 29 The non-peptidyl polymer may be linked to cysteine residue No. 30, cysteine residue 30, cysteine 31, cysteine 40, or cysteine 41, but is not particularly limited thereto.

Specifically, a reactive group of the non-peptidyl polymer may be linked to the -SH group of the cysteine residue, and all of the above descriptions apply to the reactive group. If maleimide-PEG-aldehyde is used, the maleimide group is linked to the -SH group of X by a thioether bond, and the aldehyde group is F, specifically, the -NH ₂ group of immunoglobulin Fc by reductive amination reaction may be connected through, but is not limited thereto, and this corresponds to one example.

In addition, in the conjugate, the reactive group of the non-peptidyl polymer _{may be linked to -NH 2} located at the N-terminus of the immunoglobulin Fc region, but this corresponds to one example.

Meanwhile, F may be an immunoglobulin Fc region, and more specifically, the immunoglobulin Fc region may be derived from IgG, but is not particularly limited thereto.

As used herein, the term "immunoglobulin Fc region" refers to a region including heavy chain constant region 2 (CH2) and/or heavy chain constant region 3 (CH3), excluding the heavy and light chain variable regions of immunoglobulin. The immunoglobulin Fc region may be one component constituting a moiety of the conjugate of the present invention. The immunoglobulin Fc region may be used interchangeably with “immunoglobulin Fc fragment”.

In the present specification, referring to the Fc region, not only the native sequence obtained from papain digestion of immunoglobulin, but also derivatives thereof, such as one or more amino acid residues in the native sequence, are converted by deletion, insertion, non-conservative or conservative substitution, or a combination thereof, resulting in a native natural sequence. It encompasses and includes sequences that differ from the type.

F is a structure in which two polypeptide chains are connected by a disulfide bond, and may be a structure in which only one of the two chains is connected through a nitrogen atom, but is not limited thereto. The linkage via the nitrogen atom may be linked through reductive amination to the epsilon amino atom or the N-terminal amino group of lysine.

Reductive amination reaction means a reaction in which an amine group or an amino group of a reactant reacts with an aldehyde (i.e., a functional group capable of reductive amination) of another reactant to form an amine and then forms an amine bond by a reduction reaction, It is an organic synthesis reaction well known in the art.

In one embodiment, the F may be connected through the nitrogen atom of the N-terminal proline, but is not limited thereto.

The immunoglobulin Fc region is one component constituting a moiety of the conjugate of Formula 1 of the present invention, and specifically, may correspond to F in Formula 1 above.

The immunoglobulin Fc region may include a hinge region in the heavy chain constant region, but is not limited thereto.

In the present invention, the immunoglobulin Fc region may include a specific hinge sequence at the N-terminus.

As used herein, the term “hinge sequence” refers to a region that is located on a heavy chain and forms a dimer of an immunoglobulin Fc region through an inter disulfide bond.

In the present invention, the hinge sequence may be mutated to have only one cysteine residue by deleting a part of the hinge sequence having the following amino acid sequence, but is not limited thereto:

Glu-Ser-Lys-Tyr-Gly-Pro-Pro-Cys-Pro-Ser-Cys-Pro (SEQ ID NO: 119).

The hinge sequence may include only one cysteine residue by deleting the 8th or 11th cysteine residue in the hinge sequence of SEQ ID NO: 119. The hinge sequence of the present invention may be composed of 3 to 12 amino acids, including only one cysteine residue, but is not limited thereto. More specifically, the hinge sequence of the present invention may have the following sequences: Glu-Ser-Lys-Tyr-Gly-Pro-Pro-Pro-Ser-Cys-Pro (SEQ ID NO: 120), Glu-Ser- Lys-Tyr-Gly-Pro-Pro-Cys-Pro-Ser-Pro (SEQ ID NO: 121), Glu-Ser-Lys-Tyr-Gly-Pro-Pro-Cys-Pro-Ser (SEQ ID NO: 122), Glu- Ser-Lys-Tyr-Gly-Pro-Pro-Cys-Pro-Pro (SEQ ID NO: 123), Lys-Tyr-Gly-Pro-Pro-Cys-Pro-Ser (SEQ ID NO: 124), Glu-Ser-Lys- Tyr-Gly-Pro-Pro-Cys (SEQ ID NO: 125), Glu-Lys-Tyr-Gly-Pro-Pro-Cys (SEQ ID NO: 126), Glu-Ser-Pro-Ser-Cys-Pro (SEQ ID NO: 127) , Glu-Pro-Ser-Cys-Pro (SEQ ID NO: 128), Pro-Ser-Cys-Pro (SEQ ID NO: 129), Glu-Ser-Lys-Tyr-Gly-Pro-Pro-Ser-Cys-Pro (SEQ ID NO: 129) No. 130), Lys-Tyr-Gly-Pro-Pro-Pro-Ser-Cys-Pro (SEQ ID NO: 131), Glu-Ser-Lys-Tyr-Gly-Pro-Ser-Cys-Pro (SEQ ID NO: 132), Glu-Ser-Lys-Tyr-Gly-Pro-Pro-Cys (SEQ ID NO: 133), Lys-Tyr-Gly-Pro-Pro-Cys-Pro (SEQ ID NO: 134), Glu-Ser-Lys-Pro-Ser- Cys-Pro (SEQ ID NO: 135), Glu-Ser-Pro-Ser-Cys-Pro (SEQ ID NO: 136), Glu-Pro-Ser-Cys (SEQ ID NO: 137), Ser-Cys-Pro (SEQ ID NO: 138).

More specifically, the hinge sequence may include the amino acid sequence of SEQ ID NO: 129 (Pro-Ser-Cys-Pro) or SEQ ID NO: 138 (Ser-Cys-Pro), but is not limited thereto.

The immunoglobulin Fc region of the present invention may have a form in which two molecules of an immunoglobulin Fc chain form a dimer due to the presence of a hinge sequence, and in the conjugate of Formula 1, one end of the linker is a dimer immunoglobulin It may be in a form linked to one chain of the Fc region, but is not limited thereto.

As used herein, the term "N-terminus" refers to the amino terminus of a protein or polypeptide, and 1, 2, 3, 4, 5, 6, It may include up to 7, 8, 9, or 10 or more amino acids. The immunoglobulin Fc region of the present invention may include a hinge sequence at the N-terminus, but is not limited thereto.

In addition, as long as the immunoglobulin Fc region of the present invention has substantially the same or improved effect as the native type, part or all of the heavy chain constant region 1 (CH1) and/or light chain constant region except for only the heavy and light chain variable regions of immunoglobulin 1 (CL1) may be an extended Fc region. It may also be a region in which some fairly long amino acid sequences corresponding to CH2 and/or CH3 have been removed.

For example, the immunoglobulin Fc region of the present invention comprises 1) a CH1 domain, a CH2 domain, a CH3 domain and a CH4 domain, 2) a CH1 domain and a CH2 domain, 3) a CH1 domain and a CH3 domain, 4) a CH2 domain and a CH3 domain, 5) a combination of one or two or more of the CH1 domain, the CH2 domain, the CH3 domain and the CH4 domain with an immunoglobulin hinge region (or a part of the hinge region), 6) heavy chain constant region each domain and the light chain constant region may be a dimer . However, it is not limited thereto.

In addition, in one embodiment of the long-acting conjugate of the present invention, the immunoglobulin Fc region F is a dimer (dimer) consisting of two polypeptide chains, wherein the Fc region dimers F and X are ethylene glycol repeats They are covalently linked through one and the same linker L containing the units. In one embodiment of this embodiment, X is covalently linked via a linker L to only one of the two polypeptide chains of this Fc region dimer F. In a more specific example of this embodiment, only one molecule of X is covalently linked via L to one of the two polypeptide chains of the Fc region dimer F to which X is linked. In the most specific example of this embodiment, F is a homodimer.

In another embodiment of the long-acting conjugate of the present invention, it is also possible for two molecules of X to bind symmetrically to one Fc region in a dimeric form. In this case, the immunoglobulin Fc and X may be linked to each other by a non-peptide linker. However, it is not limited to the examples described above.

In addition, the immunoglobulin Fc region of the present invention includes a native amino acid sequence as well as a sequence derivative thereof. An amino acid sequence derivative means that one or more amino acid residues in a native amino acid sequence have a different sequence by deletion, insertion, non-conservative or conservative substitution, or a combination thereof.

For example, in the case of IgG Fc, amino acid residues 214 to 238, 297 to 299, 318 to 322, or 327 to 331 known to be important for binding may be used as suitable sites for modification.

In addition, various types of derivatives are possible, such as a site capable of forming a disulfide bond is removed, some amino acids at the N-terminus of native Fc are removed, or a methionine residue may be added to the N-terminus of native Fc do. In addition, in order to eliminate the effector function, the complement binding site, eg, the C1q binding site, may be removed, or the ADCC (antibody dependent cell mediated cytotoxicity) site may be removed. Techniques for preparing such immunoglobulin Fc region sequence derivatives are disclosed in International Patent Publication Nos. WO 97/34631 and International Patent Publication No. 96/32478.

Amino acid exchanges in proteins and peptides that do not entirely alter the activity of the molecule are known in the art (H.Neurath, R.L.Hill, The Proteins, Academic Press, New York, 1979). The most commonly occurring exchanges are amino acid residues Ala/Ser, Val/Ile, Asp/Glu, Thr/Ser, Ala/Gly, Ala/Thr, Ser/Asn, Ala/Val, Ser/Gly, Thy/Phe, Ala/ Exchange between Pro, Lys/Arg, Asp/Asn, Leu/Ile, Leu/Val, Ala/Glu, Asp/Gly. In some cases, the formula (phosphorylation, sulfation, acrylation, glycosylation, methylation, farnesylation, acetylation, amidation, etc.) may be modified).

The above-described Fc derivative may exhibit biological activity equivalent to that of the Fc region of the present invention, and may have increased structural stability against heat, pH, etc. of the Fc region.

In addition, the Fc region may be obtained from a native type isolated in vivo from animals such as humans, cows, goats, pigs, mice, rabbits, hamsters, rats or guinea pigs, or obtained from transformed animal cells or microorganisms. It may be recombinant or a derivative thereof. Here, the method of obtaining from the native type may be a method of obtaining whole immunoglobulins by treatment with proteolytic enzymes after isolating whole immunoglobulins from a living body of a human or animal. When treated with papain, it is cleaved into Fab and Fc, and when treated with pepsin, it is cleaved into pF'c and F(ab) ₂ . In this case, Fc or pF'c may be separated using size-exclusion chromatography or the like. In a more specific embodiment, a recombinant immunoglobulin Fc region obtained from a human-derived Fc region from a microorganism.

In addition, the immunoglobulin Fc region may be in the form of a native sugar chain, an increased sugar chain compared to the native type, a decreased sugar chain compared to the native type, or a form in which the sugar chain is removed. Conventional methods such as chemical methods, enzymatic methods, and genetic engineering methods using microorganisms may be used for the increase or decrease or removal of such immunoglobulin Fc sugar chains. Here, the immunoglobulin Fc region from which the sugar chain is removed from the Fc has significantly reduced binding to complement (c1q) and reduced or eliminated antibody-dependent cytotoxicity or complement-dependent cytotoxicity, so that unnecessary immune responses in vivo are not induced. does not In this regard, a form more suitable for the original purpose as a drug carrier will be an immunoglobulin Fc region in which sugar chains are removed or non-glycosylated.

In the present invention, "deglycosylation" refers to an Fc region from which sugars are removed by an enzyme, and aglycosylation refers to an Fc region that is not glycosylated by production in prokaryotes, in a more specific embodiment, in E. coli. .

Meanwhile, the immunoglobulin Fc region may be of human or animal origin such as cattle, goats, pigs, mice, rabbits, hamsters, rats, and guinea pigs, and in a more specific embodiment, it is of human origin.

In addition, the immunoglobulin Fc region may be an Fc region derived from IgG, IgA, IgD, IgE, or IgM, a combination thereof, or a hybrid thereof. In a more specific embodiment, it is derived from IgG or IgM, which is most abundant in human blood, and in a more specific embodiment, it is derived from IgG, which is known to enhance the half-life of ligand binding proteins. In an even more specific embodiment, the immunoglobulin Fc region is an IgG4 Fc region, and in the most specific embodiment, the immunoglobulin Fc region is a non-glycosylated Fc region derived from human IgG4, but is not limited thereto.

In addition, in one specific embodiment, the immunoglobulin Fc fragment is a fragment of human IgG4 Fc, and is a homologue in which two monomers are linked through a disulfide bond (inter-chain form) between cysteine, amino acid 3 of each monomer. It may be in the form of a dimer, wherein each monomer of the homodimer is independently an internal disulfide bond between cysteines at positions 35 and 95 and an internal disulfide bond between cysteines at positions 141 and 199, that is, two internal disulfide bonds. (intra-chain form) and/or may have. The number of amino acids of each monomer may consist of 221 amino acids, and the amino acids forming a homodimer may consist of a total of 442 amino acids, but is not limited thereto. Specifically, in the immunoglobulin Fc fragment, two monomers having the amino acid sequence of SEQ ID NO: 139 (consisting of 221 amino acids) form a homodimer through a disulfide bond between cysteine, the 3rd amino acid of each monomer, and the monomer of the homodimer may each independently form an internal disulfide bond between cysteines at positions 35 and 95 and an internal disulfide bond between cysteines at positions 141 and 199, but is not limited thereto.

Meanwhile, in the present invention, the term "combination" with respect to an immunoglobulin Fc region means that when a dimer or multimer is formed, a polypeptide encoding a single-chain immunoglobulin Fc region of the same origin binds to a single-chain polypeptide of a different origin. means to form That is, dimers or multimers can be prepared from two or more fragments selected from the group consisting of IgG Fc, IgA Fc, IgM Fc, IgD Fc and IgE Fc fragments.

In the present invention, "hybrid" is a term that means that sequences corresponding to immunoglobulin Fc fragments of two or more different origins exist in a single-chain immunoglobulin constant region. In the case of the present invention, various types of hybrids are possible. That is, a hybrid of domains consisting of 1 to 4 domains from the group consisting of CH1, CH2, CH3 and CH4 of IgG Fc, IgM Fc, IgA Fc, IgE Fc and IgD Fc is possible, and may include a hinge.

On the other hand, IgG can also be divided into subclasses of IgG1, IgG2, IgG3 and IgG4, and in the present invention, a combination thereof or hybridization thereof is also possible. Specifically, it is an IgG2 and IgG4 subclass, and most specifically, an Fc fragment of IgG4 having little effector function such as complement dependent cytotoxicity (CDC).

In addition, the above-described conjugate may have an increased duration of effect compared to native GLP-1, GIP, or glucagon, or compared to X that is not modified with F, and such a conjugate is not only in the above-described form, but also in biodegradable nano Including all of the form encapsulated in the particles, but is not limited thereto.

In addition, the above-described conjugate may have an increased duration of effect compared to native GLP-1, GIP, or glucagon, or compared to unmodified F, and such a conjugate is not only in the form described above, but also biodegradable nanoparticles Including all enclosed forms, etc.

A composition comprising the peptide (eg, the peptide itself or a long-acting conjugate to which a biocompatible material is bound) may be used for preventing or treating lung disease.

In the present invention, the term "prevention" refers to any action that inhibits or delays the onset of lung disease by administration of the peptide (eg, the peptide itself or a long-acting conjugate to which a biocompatible material is bound) or a composition comprising the same. , "treatment" refers to any action in which the symptoms of lung disease are improved or beneficial by administration of the peptide (eg, the peptide itself or a long-acting conjugate to which a biocompatible material is bound) or a composition comprising the same.

In the present invention, the term "administration" means introducing a predetermined substance to a patient by any suitable method, and the route of administration of the composition is not particularly limited thereto, but any general route through which the composition can reach an in vivo target It may be administered through, for example, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, or rectal administration. can

The use of a triple activator having activity on both glucagon, GLP-1 and GIP receptors of the present invention or a long-acting conjugate thereof reduces the number of administrations to chronic patients who have to be administered daily due to a dramatic increase in the blood half-life and the sustained effect in vivo. It has the great advantage of improving the quality of life of patients by reducing it, which is a great help in the treatment of lung diseases. Moreover, the triple activator or a long-acting conjugate thereof has an effect in preventing lung disease, such as delaying the recurrence period of symptoms of lung disease, and/or significantly reducing symptoms of lung disease, such as prevention and / or a great help in treatment.

As used herein, the term "pulmonary disease" refers to any disease in which an abnormality occurs in tissue or function of the lung, and specifically, the triple activator of the present invention or a long-acting conjugate thereof can inhibit inflammation and fibrotic reactions. , For the purposes of the present invention, the lung disease is a disease accompanying lung inflammation and fibrosis or a disease accompanying fibrosis, for example, interstitial lung disease (ILD), progressive fibrotic interstitial lung disease (Progressive Fibrosing Interstitial Lung Disease (PF-ILD), idiopathic interstitial pneumonias (IIP), Non-specific interstitial pneumonia (NSIP), pulmonary fibrosis, interstitial fibrosing interstitial lung diseases (FILD), idiopathic pulmonary fibrosis (IPF), alveolitis, pneumonia, emphysema, bronchitis, chronic obstructive pulmonary disease, may include, but are not limited to, combined pulmonary fibrosis and emphysema (CPFE), asthma, respiratory infections, and the like.

The triple activator of the present invention inhibits the activity of macrophages, a major mechanism of lung inflammation, and inhibits the differentiation of fibroblasts into myofibroblasts and the epithelial mesenchymal transition of alveolar epithelial cells, which are the main mechanisms of pulmonary fibrosis. It may show a preventive or therapeutic effect on a disease, but is not limited thereto.

Specifically, the pharmaceutical composition of the present invention is administered by (i) inhibiting macrophage activity, and/or (ii) reducing the expression of IL-1β, IL-6, IL-12, or TNF-α. It may be to suppress inflammation, but is not limited thereto. Also, although not limited thereto, the pharmaceutical composition of the present invention may have one or more of the following characteristics upon administration:

(i) inhibition of myofibroblast differentiation;

(ii) decreased expression of α-SMA, collagen1α1, or fibronectin;

(iii) inhibition of epithelial mesenchymal transition (EMT) of alveolar epithelial cells;

(iv) decreased expression of collagen1α1, or collagen1α3. Through this, the pharmaceutical composition of the present invention may be to inhibit fibrosis, but is not limited thereto.

In the inflammatory response in the lungs, macrophages of the alveoli are involved, and neutrophils are attracted by inflammatory cytokines (IL-1, IL-6, TNF-α) secreted by the macrophages, and proteases are secreted. In particular, elastase that decomposes elastin, a type of fiber that constitutes lung tissue and is involved in elasticity, is secreted. It is known that the elastin, which is an elastic tissue of the alveoli, is damaged by the elastase, and eventually the alveoli are also damaged, leading to fibrosis of the tissue. Inflammation and fibrosis in the lungs themselves become lung diseases, but the progression and deepening of inflammation and fibrosis can lead to other lung diseases, and therefore, it is important to inhibit inflammation and fibrosis in the prevention and treatment of lung diseases. is required

The characteristics of the triple activator as described above of the present invention mean the effect of inhibiting and improving lung inflammation and fibrosis, which also suggests a preventive or therapeutic effect on lung diseases accompanying inflammation or fibrosis of the lung.

As used herein, the term chronic obstructive pulmonary disease (COPD) refers to a disease showing irreversible airway obstruction, and it is known that the destruction of the lung parenchyma and pulmonary fibrosis due to chronic inflammation and damage occur. Infiltration of various inflammatory cells is observed in the airways of patients with chronic obstructive pulmonary disease, and in particular, the number of macrophages is known to increase according to the severity of the disease, suggesting that inflammation plays an important role in the pathogenesis of chronic obstructive pulmonary disease. do. When harmful substances are inhaled, innate immunity responds and epithelial cells secrete many inflammatory mediators, which act to activate alveolar macrophages and neutrophils, so control of the inflammatory response is an important factor in the treatment of chronic obstructive pulmonary disease. . In general, chronic obstructive pulmonary disease is divided into two types of emphysema and chronic bronchitis, but it is known that emphysema and chronic bronchitis coexist in many cases in actual patients.

As used herein, the term chronic bronchitis is chronic bronchitis, which causes enlargement and increase of mucous glands in the airways, or structural damage due to inflammation. As a result, the mucus layer thickens and ciliary movement decreases, blocking air flow. Respiratory diseases that cause difficulty breathing, etc.

As used herein, the term emphysema refers to a disease in which the bronchial tubes or lungs are inflamed due to various causes, thereby secreting proteases, destroying the basic skeleton of the alveoli, and damaging the vascular structure, resulting in loss of gas exchange function.

As used herein, the term alveolitis refers to a disease in which the alveoli, which are responsible for oxygen exchange, are inflamed. If macrophages accumulate in the alveoli due to mold, dust, petroleum, toluene, acetone, chemicals, etc. and the alveolar septum thickens, it can progress to alveolitis. As the alveolitis progresses, the alveoli are destroyed, scarring, hardening, and breathing. It is known to cause trouble.

As used herein, the term asthma is an inflammatory airway obstructive disease that exhibits a reversible lesion, unlike chronic obstructive pulmonary disease, and causes respiratory distress induced by airway inflammation. It is known that the airways of asthma patients secrete a lot of cytokines such as IL-4, IL-5, and IL-13, and the proliferation and thickening of bronchial smooth muscle is remarkable. As with chronic obstructive pulmonary disease, control of the inflammatory response is important in treatment.

As used herein, the term pneumonia refers to a disease in which the parenchymal tissue of the lung or the alveoli are inflamed. Bacterial infection, virus, protozoa, fungus, and chemicals are the causes, and it is a high-risk disease that can accompany various complications. It is known that the immune mechanism (typically alveolar macrophages) of the respiratory organs against the pathogen does not function properly, or the level of the pathogen exceeds the limit that can be defended by a normal immune mechanism, leading to the onset of pneumonia. It is known that patients with chronic lung diseases such as asthma, chronic obstructive pulmonary disease, emphysema, and bronchiectasis are more likely to develop pneumonia. In the present invention, pulmonary inflammation or pneumonia may include, but is not limited to, pulmonary inflammation or pneumonia caused by or accompanying other lung diseases.

As used herein, the term "respiratory infectious disease" refers to a respiratory disease caused by infection with a pathogen (virus, bacteria, fungus, etc.). Respiratory infections can be classified according to pathogens that cause them. Representative respiratory infections include respiratory viruses, bacteria, mycoplasma, fungi, and the like. Since inflammation is accompanied by a respiratory infection caused by infection with the pathogen, a therapeutic effect through improvement of inflammation can be expected.

As used herein, the term, respiratory viral infection disease refers to a respiratory disease caused by a pathogenic virus infection, which can cause severe upper respiratory tract infections from mild upper respiratory tract infections to severe lower respiratory tract infections accompanied by pneumonia and bronchitis. The infection is known to be fatal. Respiratory virus infection causes inflammation in the respiratory tract, including the lungs, and if inflammation is not suppressed for a long period of time, it leads to fibrosis, which leads to more serious lung disease. Therefore, it is important to treat respiratory viral infections while suppressing inflammation and fibrosis.

The respiratory viruses that cause respiratory viral infections include adenovirus, vaccinia virus, herpes simplex virus, parainfluenza virus, rhinovirus, chickenpox. Virus (varicella Zoster Virus), measle virus (measle virus), respiratory syncytial virus (respiratorysyncytial virus), Dengue virus (Dengue virus), human immunodeficiency virus (HIV), influenza virus, coronavirus (coronavirus), severe acute respiratory syndrome virus (severe acute respiratory syndrome associated virus; SARS-associated virus), or middle east respiratory syndrome coronavirus (MERS-CoV), but is not limited thereto.

One non-limiting example of the coronavirus may be SARS-CoV-2, and SARS-CoV-2 infection may cause coronavirus disease 2019 (COVID-19).

In the present invention, the term, coronavirus disease 2019 (COVID-19) is a viral infectious disease caused by a coronavirus (2019-nCoV or SARS-CoV-2) infection, and the clear source and route of infection are still confirmed However, the spread was so strong that it caused a worldwide pandemic. Corona virus is a virus that can infect humans and various animals. It is an RNA virus with a gene size of 27 to 32 kb and is known to mainly exhibit respiratory symptoms such as cough with fever, shortness of breath, shortness of breath, and sputum.

In particular, the reason that severe pneumonia accompanies COVID-19 is that the coronavirus attacks bronchial ciliary epithelial cells or Type II alveolar epithelial cells (type 2 epithelial cells in the alveoli). These cells have a large amount of enzyme receptors that make the corona virus stick well. Receptors such as 'ACE2' and 'TMPRSS2' enhance the ability of corona virus to penetrate cells.

Foreign substances or pathogens that have penetrated during respiration attach to the mucous membrane of the ciliary epithelial cells of the bronchi. Ciliated epithelial cells, as their name suggests, have a large number of cilia, and they discharge pathogens including coronaviruses attached to the mucous membrane in the direction of the mouth and nose. However, there is a limit to generating many viruses at once by ciliary movement of ciliated epithelial cells. Smoking, dust, dry weather, and low temperatures also reduce ciliary motility. In dry, cold winters, ciliary movement is lowered, so influenza viral infections, including coronavirus infection-19, increase.

Like other viruses, coronaviruses seize the host cell's resources and systems, proliferate vigorously, and are ejected out of infected cells. At this time, the exponentially proliferated viruses escape and rapidly infiltrate into surrounding healthy ciliated epithelial cells and Type II alveolar epithelial cells. Infected cells secrete cytokines that cause strong inflammation and turn into inflammatory cells.

The original function of Type II alveolar epithelial cells is to secrete surfactant to keep the alveoli taut and to facilitate gas exchange through Type I alveolar epithelial cells. Type II alveolar epithelial cells attacked by the coronavirus transform into inflammatory cells and lose their main function, causing inflammation (pneumonia) in the lungs, resulting in secondary symptoms (fever, cough, shortness of breath, etc.).

The triple activator of the present invention can inhibit the inflammatory response by inhibiting macrophage activity and/or reducing the expression level of inflammatory cytokines (IL-1β, IL-6, IL-12, or TNF-α) in lung tissue. Therefore, it may exhibit a preventive or therapeutic effect on pulmonary diseases caused by or accompanied by inflammation (eg, alveolitis, pneumonia, emphysema, bronchitis, chronic obstructive pulmonary disease, asthma, or respiratory infection diseases).

On the other hand, fibrosis is a disease that forms excessive fibrous connective tissue in an organ or tissue. Fibrosis refers to a state in which normal control is impossible during the wound healing process after a tissue in the human body is damaged by an inflammatory reaction caused by various causes (infection, chemical stimulation, radiation, etc.). In particular, in the lungs, inflammation of the lungs occurs and lasts for a long time in an untreated state, leading to fibrosis that is hardened without tissue regeneration, leading to serious lung disease in many cases. Accordingly, since it is known that it is necessary to suppress inflammation to prevent the progression of fibrosis in the treatment of fibrosis, immunosuppressive agents (eg, steroids, cytoxane) and the like are used as therapeutic agents. Fibrosis may be used interchangeably with fibrosis herein.

As used herein, the term pulmonary fibrosis refers to a condition in which proliferation of fibrous connective tissue occurs in the lungs, resulting in destruction of normal lung structures and hardening and devastation of lung tissue. There are parenchymal, interstitial and mixed types, but interstitial pulmonary fibrosis is particularly problematic, in which proliferation of fibrous connective tissue appears around the alveolar wall and bronchioles. In general, transforming growth factor-β (TGF-β) is produced in various cells such as alveolar macrophages, activated alveolar epithelial cells, fibroblasts, and myofibroblasts, and fibroblast proliferation and macrophages and fibroblasts It is known to further enhance the fibrotic response by inducing the migration of inflammatory and fibrotic cytokines such as TNF-α, PDGF, IL-1β, and IL-13 (Proc Am Thorac Soc., 9( 3):111-116 (2012)). Pulmonary fibrosis of the present invention may include, but is not limited to, pulmonary fibrosis caused by or accompanying other lung diseases.

As used herein, the term combined pulmonary fibrosis and emphysema (CPFE) is a representative disease in which emphysema coexists with fibrosis.

As used herein, the term, interstitial lung disease (ILD) is also known as diffuse parenchymal lung disease (DPLD), proliferation of the lung interstitial compartment, infiltration of inflammatory cells and It is a generic term for diseases that are accompanied by fibrosis and show abnormal collagen deposition. The interstitial lung disease is classified into occupational, environmental, anthropogenic, connective tissue disease, or idiopathic according to the cause.

As used herein, the term, Progressive Fibrosing Interstitial Lung Disease (PF-ILD) refers to chronic fibrotic interstitial lung disease exhibiting a progressive phenotype, autoimmune interstitial lung disease, total sclerosis-associated interstitial lung disease, mixed connective tissue disease-associated interstitial lung disease, non-specific idiopathic interstitial pneumonia, unclassified idiopathic interstitial pneumonia, etc., but is not limited thereto.

Among the interstitial lung diseases, idiopathic interstitial pneumonia (IIP), whose cause is unknown, refers to lung diseases classified into histological forms that invade the lung interstitium, and is representative of non-specific interstitial pneumonia (Non- specific interstitial pneumonia (NSIP), and idiopathic pulmonary fibrosis (IPF).

As used herein, the term idiopathic pulmonary fibrosis (IPF) is the most common disease among idiopathic interstitial pneumonia, and is defined as pulmonary fibrosis for which the exact cause is not identified. After alveolar epithelial cells are damaged by repeated inflammatory reactions by various exposures, fibrosis is induced without normal wound healing, and pulmonary fibrosis is caused by complex interactions between genetic predisposition, environmental factors, and lung infection. known to go on. Specifically, proliferation of fibroblasts/myofibroblasts in the lungs by various factors secreted from damaged alveolar epithelial cells and infiltrated inflammatory cells, secretion and accumulation of collagen, and extracellular matrix (ECM) Excessive deposition is known as the pathogenesis.

The triple activator of the present invention inhibits (i) myofibroblast differentiation; (ii) decreased expression of α-SMA, collagen1α1, or fibronectin; (iii) inhibition of epithelial mesenchymal transition (EMT) of alveolar epithelial cells; and (iv) reduced expression of collagen1α1 or collagen1α3, so it can inhibit and improve fibrosis, and lung diseases caused by or accompanied by fibrosis (eg, interstitial lung disease, progressive fibrotic epilepsy) It may have a preventive or therapeutic effect in sexual pulmonary disease, idiopathic interstitial pneumonia, non-specific interstitial pneumonia, pulmonary fibrosis, interstitial pulmonary fibrosis, idiopathic pulmonary fibrosis, complex pulmonary fibrosis and emphysema).

On the other hand, the triple activator according to the present invention can exhibit a therapeutic effect on coronavirus infection-19, and it is known that the pulmonary fibrosis symptom remains as a sequelae after being cured of coronavirus infection-19, and the triple activator of the present invention shows an effect of improving pulmonary fibrosis, so the triple activator may exhibit efficacy against pulmonary inflammation and/or pulmonary fibrosis caused by coronavirus infection-19.

The pharmaceutical composition of the present invention may be administered in addition to mucolytic agents or a pharmaceutically acceptable salt thereof, but is not limited thereto. For the purposes of the present invention, the pharmaceutical composition is for co-administration of the triple active agent and the mucolytic agent, and the triple active agent and the mucolytic agent may be administered simultaneously, sequentially, or in reverse order, but is not limited thereto.

As used herein, the term "combination" is to be understood as indicating simultaneous, separate or sequential administration. If the administration is sequential or separate, the interval between administration of the second component should be such that the beneficial effect of the combination is not lost. The triple active agent and the mucolytic agent may be administered in combination as follows, but is not limited thereto:

a) (i) a triple active agent or a conjugate thereof and (ii) a mucolytic agent or a pharmaceutically acceptable salt thereof are administered as a mixture; or

b) (i) the triple activator or a conjugate thereof and (ii) a mucolytic agent or a pharmaceutically acceptable salt thereof are administered in an isolated form.

When the triple active agent and the mucolytic agent or a pharmaceutically acceptable salt thereof are in separate forms, the triple active agent and the mucolytic agent or a pharmaceutically acceptable salt thereof are formulated into separate formulations simultaneously, separately, sequentially, or in reverse order It may be administered as

In the present invention, combined administration does not only mean simultaneous administration, but also the triple active agent and the mucolytic agent or a pharmaceutically acceptable salt thereof act together on the subject so that each substance has a level equal to or higher than its original function. It should be understood as a dosage form capable of carrying out

As a specific example, the triple active agent and the mucolytic agent or a pharmaceutically acceptable salt thereof may be mixed and administered together as one formulation, or the triple active agent and the mucolytic agent or a pharmaceutically acceptable salt thereof may be separately administered. It may be formulated to be administered concurrently, sequentially, or in reverse order, but is not limited thereto. When formulated separately and administered in combination, each formulation may be administered by different routes, but is not limited thereto. In addition, although not limited thereto, the triple active agent and the mucolytic agent or a pharmaceutically acceptable salt thereof may be separately formulated and included in one kit. Pharmaceutically acceptable salts of mucolytic agents include salts derived from pharmaceutically acceptable inorganic acids, organic acids, or bases. Examples of suitable acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid , benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include alkali metals such as sodium and potassium, alkaline earth metals such as magnesium, and ammonium.

As used herein, the term “mucolytic agents” refers to drugs that promote the secretion, liquefaction, or discharge of sputum, sputum or mucus from the respiratory tract, in particular, until the mucus in the lungs is decomposed to dilute the respiratory secretions. A drug that only works. In the present invention, the mucolytic agent may be used in combination with an expectorant.

Specific examples of the mucolytic agent of the present invention include ambroxol, N-acetylcysteine, N-acetylin, carbocysteine, domiodol, fudosteine, bromhexine, erdosteine, letostine, lysozyme, mesna, sobrerol, stepronin, thio Pronin, tyloxapol, carbocisteine, dornase alfa, eprazinone, letosteine, neltenexine, and mecysteine ) may be any one or more selected from the group consisting of, but is not limited thereto.

On the other hand, the combined administration of the triple activator and the mucolytic agent may exhibit a therapeutic effect on respiratory infectious diseases (eg, coronavirus infection-19). Since the triple activator of the present invention exhibits an effect of improving lung inflammation and pulmonary fibrosis, the combined administration of the triple activator and a mucolytic agent is a respiratory infection disease (eg, coronavirus infection-19 (COVID-19)), or It may show efficacy against pulmonary inflammation and pulmonary fibrosis.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier or diluent. Such pharmaceutically acceptable carriers, or diluents, may be non-naturally occurring.

In the present invention, the term "pharmaceutically acceptable" means a sufficient amount to exhibit a therapeutic effect and does not cause side effects, and the type of disease, the patient's age, weight, health, sex, and the patient's sensitivity to the drug , administration route, administration method, frequency of administration, treatment period, combination or drugs used at the same time can be easily determined by those skilled in the art according to factors well known in the medical field.

The pharmaceutical composition including the peptide of the present invention may include a pharmaceutically acceptable excipient. The excipient is not particularly limited thereto, but in the case of oral administration, a binder, a lubricant, a disintegrant, a solubilizer, a dispersing agent, a stabilizer, a suspending agent, a color, a fragrance, etc. may be used, and in the case of an injection, a buffer, a preservative, An analgesic agent, a solubilizer, an isotonic agent, a stabilizer, etc. may be mixed and used, and in the case of topical administration, a base, excipient, lubricant, preservative, etc. may be used.

The formulation of the composition of the present invention can be prepared in various ways by mixing with the pharmaceutically acceptable excipients as described above. For example, in the case of oral administration, it may be prepared in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like, and in the case of injections, it may be prepared in the form of unit dose ampoules or multiple doses. In addition, it can be formulated as a solution, suspension, tablet, pill, capsule, sustained-release preparation, and the like.

Meanwhile, examples of suitable carriers, excipients and diluents for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil may be used. In addition, it may further include a filler, an anti-aggregating agent, a lubricant, a wetting agent, a flavoring agent, a preservative, and the like.

In addition, the pharmaceutical composition of the present invention is any one selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, internal solutions, emulsions, syrups, sterile aqueous solutions, non-aqueous solvents, freeze-dried preparations and suppositories may have the form of

In addition, the composition is formulated in a dosage form suitable for administration in a patient's body according to a conventional method in the pharmaceutical field, specifically, a formulation useful for administration of a protein drug, and administration commonly used in the art. Oral, or dermal, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intraventricular, pulmonary, transdermal, subcutaneous, intraperitoneal, intranasal, gastrointestinal, topical, sublingual, intravaginal or It may be administered by parenteral routes of administration including, but not limited to, rectal routes.

In addition, the conjugate can be used by mixing with various pharmaceutically acceptable carriers such as physiological saline or organic solvents, and carbohydrates such as glucose, sucrose or dextran, ascorbic acid (ascorbic acid) in order to increase stability or absorption. acid) or antioxidants such as glutathione, chelating agents, low molecular weight proteins or other stabilizers, etc. may be used as agents.

In another aspect of the present invention, the prevention of lung disease comprising administering to an individual a pharmaceutical composition containing a peptide comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 102 or a long-acting conjugate thereof in a pharmaceutically effective amount treatment methods are provided.

The dosage and frequency of administration of the pharmaceutical composition of the present invention is determined according to the type of drug as the active ingredient, along with several related factors such as the disease to be treated, the route of administration, the age, sex and weight of the patient, and the severity of the disease. Specifically, the composition of the present invention may include, but is not limited to, the triple active agent or a long-acting conjugate including the same in a pharmaceutically effective amount.

The inclusion of the peptide or the long-acting conjugate in a pharmaceutically effective amount means the degree to which the desired pharmacological activity (for example, prevention, improvement or treatment of lung disease) can be obtained due to the triple activator or the long-acting conjugate, and , It may also mean a pharmaceutically acceptable level as a level that does not cause toxicity or side effects in the administered subject or is insignificant, but is not limited thereto. Such a pharmaceutically effective amount may be determined by comprehensively considering the number of administration, patient, formulation, and the like.

Although not particularly limited thereto, the pharmaceutical composition of the present invention may contain the component (active ingredient) in an amount of 0.01 to 99% by weight to volume.

The total effective amount of the composition of the present invention may be administered to a patient as a single dose, and may be administered by a fractionated treatment protocol in which multiple doses are administered for a long period of time. The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the severity of the disease. Specifically, the preferred total dose of the triple active agent or long-acting conjugate thereof of the present invention may be about 0.0001 mg to 500 mg per kg of body weight of the patient per day. However, the dose of the triple activator or its conjugate is determined by considering various factors such as the age, weight, health status, sex, severity of disease, diet and excretion rate of the patient as well as the administration route and number of treatments of the pharmaceutical composition. Since the effective dosage is determined, those of ordinary skill in the art will be able to determine an appropriate effective dosage according to the specific use of the composition of the present invention in consideration of this point. The pharmaceutical composition according to the present invention is not particularly limited in its formulation, administration route and administration method as long as the effect of the present invention is exhibited.

The pharmaceutical composition of the present invention has excellent in vivo persistence and potency, and can significantly reduce the number and frequency of administration of the pharmaceutical preparation of the present invention.

Another aspect embodying the present invention is the triple activator (peptide) and / or a long-acting conjugate of the triple activator, or a composition comprising the same, comprising administering to an individual in need thereof, lung disease A method of preventing or treating is provided.

The triple activator and/or a long-acting conjugate of the triple activator, or a composition comprising the same, lung disease, prevention and treatment are the same as described above.

In the present invention, the subject is a subject suspected of lung disease, and the subject suspected of lung disease refers to mammals including mice, livestock, etc. including humans that have or may develop the disease, but the triple activity of the present invention Subjects that can be treated with the body and/or conjugate, or the composition comprising the same, are included without limitation. In particular, it may be an individual having inflammation and/or fibrosis in the lung, or an individual having a pulmonary disease caused by or accompanied by inflammation and/or fibrosis, but is not limited thereto.

In the present invention, the term "administration" means introducing a predetermined substance to a patient by any suitable method, and the route of administration of the composition is not particularly limited thereto, but any general route through which the composition can reach an in vivo target It may be administered through, for example, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, or rectal administration. can

The method of the present invention may include administering a pharmaceutical composition comprising the triple active agent or a long-acting conjugate thereof in a pharmaceutically effective amount. A suitable total daily amount may be determined by the treating physician within the scope of sound medical judgment, and may be administered once or divided into several doses. However, for the purposes of the present invention, a specific therapeutically effective amount for a particular patient depends on the type and extent of the response to be achieved, the specific composition, including whether other agents are used, if necessary, the specific composition, the patient's age, weight, general health, It is preferable to apply differently depending on various factors including sex and diet, administration time, administration route and secretion rate of the composition, treatment period, drugs used together or concurrently with a specific composition, and similar factors well known in the pharmaceutical field.

Although not limited thereto, the pharmaceutical composition of the present invention may be administered once a week, once every 2 weeks, or once every 4 weeks, but is not limited thereto.

As a specific example of the method for preventing or treating lung disease of the present invention, a triple active agent (peptide) and/or a long-acting conjugate of the triple active agent, or a composition comprising the same, and a mucolytic agent or a pharmaceutically acceptable salt thereof are simultaneously administered. , sequentially, or in the reverse order, but is not limited thereto. The combined administration is the same as described above.

Another embodiment embodying the present invention is the use of a composition comprising the triple active agent or a long-acting conjugate thereof in the manufacture of a medicament for the prevention or treatment of lung disease.

The triple activator and/or its conjugate, or a composition comprising the same, lung disease, prevention and treatment are the same as described above.

Another aspect embodying the present invention is to provide the use of the triple active agent or a long-acting conjugate thereof, or a composition comprising the same for the prevention or treatment of lung disease.

The triple activator and/or its conjugate, or a composition comprising the same, lung disease, prevention and treatment are the same as described above.

The use of the present invention for the prevention or treatment of lung diseases is for the combined administration of a triple active agent (peptide) and/or a long-acting conjugate of the triple active agent, or a composition comprising the same, and a mucolytic agent or a pharmaceutically acceptable salt thereof. may be used, but is not limited thereto. The combined administration is the same as described above.

Hereinafter, the present invention will be described in more detail by way of Examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Example 1: Preparation of triple activator

A triple activator exhibiting activity on both GLP-1, GIP and glucagon receptors was prepared, and its sequences are shown in Table 1 below.

SEQ ID NO: order Information One H X Q G T F T S D V S S Y L D G Q A A K E F I A W L V K G C 2 H X Q G T F T S D V S S Y L D G Q A Q K E F I A W L V K G C 3 H X Q G T F T S D V S S Y L L G Q A A K Q F I A W L V K G G G P S S G A P P P S C 4 H X Q G T F T S D V S S Y L L G Q Q Q K E F I A W L V K G C 5 H X Q G T F T S D V S S Y L L G Q Q Q K E F I A W L V K G G G P S S G A P P P S C 6 H X Q G T F T S D V S S Y L D G Q A A K E F V A W L L K G C 7 H X Q G T F T S D V S K Y L D G Q A A K E F V A W L L K G C 8 H X Q G T F T S D V S K Y L D G Q A A Q E F V A W L L K G C 9 H X Q G T F T S D V S K Y L D G Q A A Q E F V A W L L A G C 10 H X Q G T F T S D V S K Y L D G Q A A Q E F V A W L L A G G G P S S G A P P P S C 11 CA G E G T F T S D L S K Y L D S R R Q Q L F V Q W L K A G G P S S G A P P P S H G 12 CA G E G T F I S D L S K Y M D E Q A V Q L F V E W L M A G G P S S G A P P P S H G 13 CA G E G T F I S D Y S I Q L D E I A V Q D F V E W L L A Q K P S S G A P P P S H G 14 CA G Q G T F T S D Y S I Q L D E I A V R D F V E W L K N G G P S S G A P P P S H G 15 CA G Q G T F T S D L S K Q M D E E A V R L F I E W L K N G G P S S G A P P P S H G 16 CA G Q G T F T S D L S K Q M D S E A Q Q L F I E W L K N G G P S S G A P P P S H G 17 CA G Q G T F T S D L S K Q M D E E R A R E F I E W L L A Q K P S S G A P P P S H G 18 CA G Q G T F T S D L S K Q M D S E R A R E F I E W L K N T G P S S G A P P P S H G 19 CA G Q G T F T S D L S I Q Y D S E H Q R D F I E W L K D T G P S S G A P P P S H G 20 CA G Q G T F T S D L S I Q Y E E E A Q Q D F V E W L K D T G P S S G A P P P S H G 21 YXQGTFTSDYSKYLD E CRA K EFVQWLLDHHPSSGQPPPS ring formation 22 YXQGTFTSDYSKCLD E KRA K EFVQWLLDHHPSSGQPPPS ring formation 23 YXQGTFTSDYSKYLD E CRA K EFVQWLLAQKGKKKNDWKHNIT ring formation 24 YXQGTFTSDYSKYLD E CRA K EFVQWLKNGGPSSGAPPPS ring formation 25 H X Q G T F T S D C S K Y L D E R A A Q D F V Q W L L D G G P S S G A P P P S 26 H X Q G T F T S D C S K Y L D S R A A Q D F V Q W L L D G G P S S G A P P P S 27 H X Q G T F T S D Y S K Y L D E R A C Q D F V Q W L L D Q G G P S S G A P P P S 28 H X Q G T F T S D Y S K Y L D E K R A Q E F V C W L L A Q K G K K N D W K H N I T 29 HXQGTFTSDYSKYLD E KAA K EFVQWLLNTC ring formation 30 HXQGTFTSDYSKYLD E KAQ K EFVQWLLDTC ring formation 31 HXQGTFTSDYSKYLD E KAC K EFVQWLLAQ ring formation 32 HXQGTFTSDYSKYLD E KAC K DFVQWLLDGGPSSGAPPPS ring formation 33 HXQGTFTSDYSIAMD E IHQ K DFVNWLLAQKC ring formation 34 HXQGTFTSDYSKYLD E KRQ K EFVNWLLAQKC ring formation 35 HXQGTFTSDYSIAMD E IHQ K DFVNWLLNTKC ring formation 36 HXQGTFTSDYSKYLC E KRQ K EFVQWLLNGGPSSGAPPPSG ring formation 37 HXQGTFTSDYSKYLD E CRQ K EFVQWLLNGGPSSGAPPPSG ring formation 38 CA X Q G T F T S D K S S Y L D E R A A Q D F V Q W L L D G G P S S G A P P P S S 39 H X Q G T F T S D Y S K Y L D G Q H A Q C F V A W L L A G G G P S S G A P P P S 40 H X Q G T F T S D K S K Y L D E R A C Q D F V Q W L L D G G P S S G A P P P S 41 H X Q G T F T S D K S K Y L D E C A A Q D F V Q W L L D G G P S S G A P P P S 42 YXQGTFTSDYSKYLD E KRA K EFVQWLLDHHPSSGQPPPSC ring formation 43 YXQGTFTSDYSKYLD E KRA K EFVQWLLDHHCSSGQPPPS ring formation 44 H G Q G T F T S D C S K Q L D G Q A A Q E F V A W L L A G G P S S G A P P P S 45 H G Q G T F T S D C S K Y M D G Q A A Q D F V A W L L A G G P S S G A P P P S 46 H G Q G T F T S D C S K Y L D E Q H A Q E F V A W L L A G G P S S G A P P P S 47 H G Q G T F T S D C S K Y L D G Q R A Q E F V A W L L A G G P S S G A P P P S 48 H G Q G T F T S D C S K Y L D G Q R A Q D F V N W L L A G G P S S G A P P P S 49 CA XQGTFTSDYSICMD E IHQ K DFVNWLLNTK ring formation 50 HXQGTFTSDYSKYLD E KRA K EFVQWLLDHHPSSGQPPPSC ring formation 51 HXQGTFTSDYSKYLD E KRQ K EFVQWLLNTC ring formation 52 HXQGTFTSDYSKYLD E KRQ K EFVQWLLDTC ring formation 53 HXEGTFTSDYSIAMD E IHQ K DFVNWLLAQC ring formation 54 HXEGTFTSDYSIAMD E IHQ K DFVDWLLAEC ring formation 55 HXQGTFTSDYSIAMD E IHQ K DFVNWLLAQC ring formation 56 HXQGTFTSDYSKYLD E KRQ K EFVNWLLAQC ring formation 57 HXQGTFTSDYSIAMD E IHQ K DFVNWLLNTC ring formation 58 HXQGTFTSDYSKYLD E KRQ K EFVQWLLNTKC ring formation 59 CA XQGTFTSDYSICMD E KHQ K DFVNWLLNTK ring formation 60 CA XQGTFTSDYSIAMD E KHC K DFVNWLLNTK ring formation 61 CA XQGTFTSDYSIAMD E IAC K DFVNWLLNTK ring formation 62 CA X Q G T F T S D K S K Y L D E R A A Q D F V Q W L L D G G P S S G A P P P S 63 CA X Q G T F T S D C S K Y L D E R A A Q D F V Q W L L D G G P S S G A P P P S 64 YXQGTFTSDYSKYLD E CAA K EFVQWLLDHHPSSGQPPPS ring formation 65 HXQGTFTSDYSKCLD E KRA K EFVQWLLDHHPSSGQPPPS ring formation 66 YXQGTFTSDYSKYLD E CRA K DFVQWLLDHHPSSGQPPPS ring formation 67 YXQGTFTSDYSKYLD E CAA K DFVQWLLDHHPSSGQPPPS ring formation 68 YXQGTFTSDYSKCLD E KAA K EFVQWLLDHHPSSGQPPPS ring formation 69 YXQGTFTSDYSKCLD E RAA K EFVQWLLDHHPSSGQPPPS ring formation 70 YXQGTFTSDYSKCLD E KRA K DFVQWLLDHHPSSGQPPPS ring formation 71 YXQGTFTSDYSKYLD E RAC K DFVQWLLDHHPSSGQPPPS ring formation 72 YXQGTFTSDCSKYLD E RAA K DFVQWLLDHHPSSGQPPPS ring formation 73 CA XQGTFTSDYSKYLD E CRA K EFVQWLLDHHPSSGQPPPS ring formation 74 CA XQGTFTSDYSKCLD E KRA K EFVQWLLDHHPSSGQPPPS ring formation 75 YXQGTFTSDYSKYLD E KAA K EFVQWLLDHHPSSGQPPPSC ring formation 76 YXQGTFTSDYSKYLD E KRA K DFVQWLLDHHPSSGQPPPSC ring formation 77 YXQGTFTSDYSKYLD E KAA K DFVQWLLDHHPSSGQPPPSC ring formation 78 HXQGTFTSDYSKYLD E KRQ K EFVQWLLDTKC ring formation 79 HXEGTFTSDYSIAMD E IHQ K DFVNWLLAQKC ring formation 80 HXEGTFTSDYSIAMD E IHQ K DFVDWLLAEKC ring formation 81 CA XQGTFTSDYSKYLD E KRQ K EFVQWLLNTC ring formation 82 CA XQGTFTSDYSKYLD E KRQ K EFVQWLLDTC ring formation 83 CA XEGTFTSDYSIAMD E IHQ K DFVNWLLAQC ring formation 84 CA XEGTFTSDYSIAMD E IHQ K DFVDWLLAEC ring formation 85 CA XQGTFTSDYSIAMD E IHQ K DFVNWLLAQC ring formation 86 CA XQGTFTSDYSKYLD E KRQ K EFVNWLLAQC ring formation 87 CA XQGTFTSDYSIAMD E IHQ K DFVNWLLNTC ring formation 88 CA XQGTFTSDYSKYLD E KRQ K EFVQWLLNTKC ring formation 89 CA XQGTFTSDYSKYLD E KRQ K EFVQWLLDTKC ring formation 90 CA XEGTFTSDYSIAMD E IHQ K DFVNWLLAQKC ring formation 91 CA XEGTFTSDYSIAMD E IHQ K DFVDWLLAEKC ring formation 92 CA XQGTFTSDYSIAMD E IHQ K DFVNWLLAQKC ring formation 93 CA XQGTFTSDYSKYLD E KRQ K EFVNWLLAQKC ring formation 94 CA XQGTFTSDYSIAMD E IHQ K DFVNWLLNTKC ring formation 95 YXQGTFTSDYSKYLD E KRA K EFVQWLLCHHPSSGQPPPS ring formation 96 YXQGTFTSDYSKYLD E KRA K EFVQWLLDHCPSSGQPPPS ring formation 97 YXQGTFTSDYSKYLD E KRA K EFVQWLLDCHPSSGQPPPS ring formation 98 YXQGTFTSDYSKALD E KAA K EFVNWLLDHHPSSGQPPPSC ring formation 99 YXQGTFTSDYSKALD E KAA K DFVNWLLDHHPSSGQPPPSC ring formation 100 YXQGTFTSDYSKALD E KAA K EFVQWLLDQHPSSGQPPPSC ring formation 101 YXQGTFTSDYSKALD E KAA K EFVNWLLDQHPSSGQPPPSC ring formation 102 YXQGTFTSDYSKALD E KAA K DFVNWLLDQHPSSGQPPPSC ring formation

In the sequence shown in Table 1, an amino acid denoted by X is a non-natural amino acid Aib (2-aminoisobutyric acid), and an underlined amino acid means that the underlined amino acids form a ring with each other. In addition, in Table 1, CA means 4-imidazoacetyl (4-imidazoacetyl), Y means tyrosine.

Example 2: Preparation of long-acting conjugates of triple activators

10 kDa PEG, that is, maleimide-PEG-aldehyde (10 kDa, NOF, Japan) having a maleimide group and an aldehyde group at both ends, respectively, was added to the triple activator of Example 1 (SEQ ID NOs: 21, 22, 42, 43, 50) , 77, and 96) for pegylation to the cysteine residues, the molar ratio of the triple activator to maleimide-PEG-aldehyde is 1:1 to 3, and the protein concentration is 1 to 5 mg/ml, and 0.5 to The reaction was carried out for 3 hours. At this time, the reaction was performed under an environment in which 20 to 60% isopropanol was added to 50 mM Tris buffer (pH 7.5). After completion of the reaction, the reaction solution was applied to SP Sepharose HP (GE healthcare, USA) to purify the tri-activator mono-pegylated to cysteine.

Next, the purified mono-pegylated tri-activator and immunoglobulin Fc (homodimer of SEQ ID NO: 139) were mixed at a molar ratio of 1:1 to 5 and a protein concentration of 10 to 50 mg/ml at 4 to 8°C. reacted for 12 to 18 hours. The reaction was carried out in an environment in which 10 to 50 mM sodium cyanoborohydride and 10 to 30% isopropanol as reducing agents were added to 100 mM potassium phosphate buffer (pH 6.0). After completion of the reaction, the reaction solution was applied to a butyl sepharose FF purification column (GE healthcare, USA) and a Source ISO purification column (GE healthcare, USA) to purify a conjugate containing the triple activator and immunoglobulin Fc. did. This purified long-acting conjugate has a structure in which a triple active peptide, a polyethylene glycol (PEG) linker and an Fc dimer are covalently linked in a molar ratio of 1:1:1, and the PEG linker is one of the two polypeptide chains of the Fc dimer. connected to only one chain.

On the other hand, in the immunoglobulin Fc, two monomers having the amino acid sequence of SEQ ID NO: 139 (consisting of 221 amino acids) form a homodimer through a disulfide bond between cysteine, which is the 3rd amino acid of each monomer, and the monomer of the homodimer Each independently formed an internal disulfide bond between cysteines at positions 35 and 95 and an internal disulfide bond between cysteines at positions 141 and 199.

After preparation, the purity analyzed by reverse-phase chromatography, size exclusion chromatography, and ion exchange chromatography was 95% or more.

Herein, a conjugate in which the triple activator of SEQ ID NO: 21 and immunoglobulin Fc are linked through a PEG linker was termed a 'conjugate comprising SEQ ID NO: 21 and immunoglobulin Fc' or a 'continuous conjugate of SEQ ID NO: 21'. It may be used interchangeably herein.

Here, the conjugate in which the triple activator of SEQ ID NO: 22 and the immunoglobulin Fc are linked through a PEG linker was designated as a 'conjugate comprising SEQ ID NO: 22 and immunoglobulin Fc' or a 'continuous conjugate of SEQ ID NO: 22', which It may be used interchangeably herein.

Here, the conjugate in which the triple activator of SEQ ID NO: 42 and the immunoglobulin Fc are linked via PEG was designated as a 'conjugate comprising SEQ ID NO: 42 and immunoglobulin Fc' or a 'continuous conjugate of SEQ ID NO: 42'. may be used interchangeably.

Here, the conjugate in which the triple activator of SEQ ID NO: 43 and the immunoglobulin Fc are linked through PEG was designated as a 'conjugate comprising SEQ ID NO: 43 and immunoglobulin Fc' or a 'continuous conjugate of SEQ ID NO: 43'. can be used interchangeably in

Herein, a conjugate in which the triple activator of SEQ ID NO: 50 and immunoglobulin Fc are linked via PEG was designated as a 'conjugate comprising SEQ ID NO: 50 and immunoglobulin Fc' or a 'continuous conjugate of SEQ ID NO: 50'. may be used interchangeably.

Here, the conjugate in which the triple activator of SEQ ID NO: 77 and the immunoglobulin Fc are linked through PEG was designated as a 'conjugate comprising SEQ ID NO: 77 and an immunoglobulin Fc' or a 'persistent conjugate of SEQ ID NO: 77'. can be used interchangeably in

Here, the conjugate in which the triple activator of SEQ ID NO: 96 and the immunoglobulin Fc are linked through PEG was designated as a 'conjugate comprising SEQ ID NO: 96 and immunoglobulin Fc' or a 'continuous conjugate of SEQ ID NO: 96'. may be used interchangeably.

Experimental Example 1: Triple activator and its long-acting conjugate in vitro Activity measurement

In order to measure the activity of the triple activator prepared in Examples 1 and 2 and the long-acting conjugate thereof, cells were in vitro using cell lines transformed with GLP-1 receptor, glucagon (GCG) receptor, and GIP receptor, respectively. A method for measuring activity was used.

Each of the above cell lines was transformed to express human GLP-1 receptor, human GCG receptor and human GIP receptor genes in Chinese hamster ovary (CHO), respectively, and is suitable for measuring the activities of GLP-1, GCG and GIP. Therefore, the activity for each part was measured using each transformed cell line.

Human GLP-1 was serially diluted from 50 nM to 0.000048 nM by 4 times to measure the GLP-1 activity of the triple activator and its long-acting conjugate prepared in Examples 1 and 2, and in Examples 1 and 2 The prepared triple activator and its long-acting conjugate were serially diluted from 400 nM to 0.00038 nM by 4 times. The culture medium was removed from the cultured CHO cells expressing the human GLP-1 receptor, 5 μl of each serially diluted substance was added to the cells, and then 5 μl of a buffer containing cAMP antibody was added for 15 minutes. during incubation at room temperature. Then, 10 μl of a detection mix containing a cell lysis buffer was added to lyse the cells and reacted at room temperature for 90 minutes. The cell lysate from which the reaction was completed was applied to the LANCE cAMP kit (PerkinElmer, USA) _{to calculate the EC 50} value through the accumulated cAMP, and then compared with each other. Relative titers compared to human GLP-1 are shown in Tables 2 and 3 below.

Human GCG was serially diluted from 50 nM to 0.000048 nM by 4 times in order to measure the GCG activity of the triple activator and the long-acting conjugate prepared in Examples 1 and 2, and the triple activity prepared in Examples 1 and 2 The sieve and its long-acting conjugate were serially diluted from 400 nM to 0.00038 nM in 4-fold increments. The culture medium was removed from the cultured CHO cells expressing the human GCG receptor, 5 μl of each serially diluted substance was added to the cells, and 5 μl of a buffer containing cAMP antibody was added thereto, followed by room temperature for 15 minutes. cultured in Then, 10 μl of a detection mix containing cell lysis buffer was added to lyse the cells, followed by reaction at room temperature for 90 minutes. The cell lysate after the reaction was completed was applied to the LANCE cAMP kit (PerkinElmer, USA) _{to calculate the EC 50} value through the accumulated cAMP, and then compared with each other. Relative titers compared to human GCG are shown in Tables 2 and 3 below.

In order to measure the GIP activity of the triple activator prepared in Examples 1 and 2 and the long-acting conjugate thereof, human GIP was serially diluted from 50 nM to 0.000048 nM by 4 times, and the triple activity prepared in Examples 1 and 2 above. The sieve and its long-acting conjugate were serially diluted from 400 nM to 0.00038 nM in 4-fold increments. The culture medium was removed from the cultured CHO cells expressing the human GIP receptor, 5 μl of each serially diluted substance was added to the cells, and 5 μl of a buffer containing cAMP antibody was added thereto, followed by room temperature for 15 minutes. cultured in Then, 10 μl of a detection mix containing cell lysis buffer was added to lyse the cells, followed by reaction at room temperature for 90 minutes. The cell lysate after the reaction was completed was applied to the LANCE cAMP kit (PerkinElmer, USA) _{to calculate the EC 50} value through the accumulated cAMP, and then compared with each other. Relative titers compared to human GIP are shown in Tables 2 and 3 below.

Relative potency ratio of triple active substances In vitro activity compared to native peptide (%) SEQ ID NO: vs GLP-1 vs Glucagon vs GIP One 3.2 <0.1 <0.1 2 5.9 <0.1 <0.1 3 1.8 <0.1 <0.1 4 8.5 <0.1 <0.1 5 42.1 <0.1 <0.1 6 17.0 <0.1 <0.1 7 13.7 <0.1 <0.1 8 14.2 0.10 <0.1 9 32.1 0.13 <0.1 10 46.0 <0.1 <0.1 11 1.4 <0.1 <0.1 12 0.4 <0.1 <0.1 13 < 0.1 < 0.1 < 0.1 14 28.0 < 0.1 < 0.1 15 79.2 <0.1 <0.1 16 2.1 < 0.1 < 0.1 17 0.2 < 0.1 < 0.1 18 <0.1 <0.1 <0.1 19 <0.1 <0.1 <0.1 20 <0.1 <0.1 <0.1 21 17.8 267 22.7 22 20.1 140 59.7 23 4.01 9.3 <0.1 24 41.2 9.3 < 0.1 25 82.6 0.1 <0.1 26 64.5 0.2 <0.1 27 83.1 0.8 0.9 28 17.2 1.6 <0.1 29 38.5 6.0 <0.1 30 142 0.7 0.8 31 135 2.2 2.4 32 151 1.7 8.8 33 24.5 <0.1 10.4 34 19.1 0.92 0.6 35 7.5 <0.1 1.3 36 37.4 0.39 0.2 37 236 6.21 2.2 38 2.3 - - 39 13.9 0.53 <0.1 40 75.2 <0.1 <0.1 41 34.3 <0.1 <0.1 42 33.9 205.8 7.8 43 12.6 88.4 3.70 44 1.3 <0.1 <0.1 45 6.6 < 0.1 < 0.1 46 1.4 < 0.1 < 0.1 47 2.4 < 0.1 < 0.1 48 1.5 < 0.1 < 0.1 49 29.8 <0.1 3.3 50 67.4 50.5 2.7 51 14.4 2.0 0.1 52 44.1 7.5 0.3 53 161 8.4 1.3 54 30.6 1.4 0.1 55 27.1 0.7 2.4 56 57.9 4.9 0.8 57 11.7 <0.1 0.3 58 39.1 2.6 0.2 59 40.3 <0.1 4.0 60 106.2 <0.1 8.2 61 59.8 <0.1 2.8 62 5.2 <0.1 <0.1 63 15.3 <0.1 <0.1 64 64.6 60.1 92.9 65 95.4 25.2 11.6 66 15.8 172 17.2 67 28.5 46.2 39.8 68 27.9 8.8 107 69 24.3 9.6 62.8 70 15.1 71.3 64.4 71 90.1 12.7 94.7 72 11.5 1.0 1.6 73 22.6 5.4 3.0 74 12.9 0.9 1.0 75 35.1 8.5 18.0 76 10.3 47.6 11.7 77 38.7 12.2 35.5 78 51.0 14.0 0.12 79 41.5 4.9 1.4 80 8.1 0.0 0.1 81 7.8 0.3 <0.1 82 9.5 1.1 <0.1 83 47.3 1.3 0.4 84 4.2 <0.1 <0.1 85 4.3 <0.1 0.3 86 28.4 0.4 0.2 87 0.9 <0.1 <0.1 88 9.6 0.3 <0.1 89 7.1 0.7 <0.1 90 7.4 <0.1 <0.1 91 31.9 16.8 0.3 92 0.8 <0.1 0.4 93 5.7 0.3 0.7 94 0.5 <0.1 <0.1 95 2.1 0.4 <0.1 96 34.4 194.8 5.2 97 10.5 62.8 2.6 98 28.1 8.2 47.1 99 20.9 14.9 57.7 100 42.2 12.7 118.5 101 23.2 13.9 40.1 102 23.3 29.5 58.0

Relative potency ratio of triple active long-acting conjugates long-acting compound In vitro activity compared to native peptide (%) vs GLP-1 vs Glucagon vs GIP 21 0.1 1.6 0.2 22 0.1 0.9 0.5 42 3.1 23.1 1.2 43 2.1 13.5 0.6 50 15.4 6.9 0.7 77 6.7 1.7 6.6 96 0.3 4.0 0.3

The novel triple activator long-acting conjugate prepared above has a function as a triple activator capable of activating all of the GLP-1 receptor, GIP receptor and glucagon receptor, and thus can be used as a therapeutic agent for lung diseases.

Experimental Example 2: Triple activator long-acting conjugate Confirmation of the effect of improving inflammation in the lungs

Whether the triple activator long-acting conjugate according to the present invention prepared in the above Example exhibits an effect of improving inflammation in the lungs was confirmed using mice administered with elastase (ELA).

Specifically, C57BL/6 mice (ORIENT Bio, Busan, Korea) treated with ELA 0.2U/mouse were treated with an excipient control group, the long-acting conjugate of SEQ ID NO: 42 (hereinafter, triple-activator long-acting conjugate; 6.5 nmol/kg) , Q2D, subcutaneous) administration group, and anti-inflammatory and COPD treatment roflumilast (10 mg/kg, QD, oral) administration group, and repeated administration was performed for 3 weeks. Thereafter, the degree of change in the expression of inflammatory cytokines in the lung tissue according to the administration of the excipient, the triple-activator long-acting conjugate, and roflumilast was confirmed through qPCR. Statistical treatment evaluated the efficacy of triple-activator long-acting conjugates using one-way ANOVA (* ~ ** p <0.05 ~ 0.01).

As a result, when the triple activator long-acting conjugate was repeatedly administered, the expression of IL-1β, IL-6, IL-12, and TNF-α in the lung tissue was significantly and equal to or greater than that of the excipient control group and the roflumilast administration group, respectively. It was confirmed that there was a decrease (Fig. 1).

Through this, it was confirmed that the triple activator long-acting conjugate of the present invention has the effect of suppressing and improving inflammation in the lungs.

Experimental Example 3: Triple activator long-acting conjugate Confirmation of treatment effect for emphysema and chronic obstructive pulmonary disease

The present inventors tried to confirm whether the triple activator long-acting conjugate could exhibit in vivo therapeutic effects on emphysema and chronic obstructive pulmonary disease (COPD), which are typical lung diseases and diseases caused by lung inflammation.

For this purpose, an animal model in which chronic obstructive pulmonary disease was induced by damage to lung tissue by intratracheal administration of elastase was used. Specifically, C57BL/6 mice (ORIENT Bio, Busan, Korea) treated with elastase at 0.2 U/meat were treated with COPD mice as excipient control group, triple activator long-acting conjugate (6.5 nmol/kg, Q2D, subcutaneous) administration group. , was divided into roflumilast (10 mg/kg, QD, oral) administration group, and repeated administration was performed for 3 weeks. After repeated administration for 3 weeks, the lung tissue of each mouse was taken as an autopsy, and the degree of emphysema of the lung tissue was evaluated through H&E staining.

As a result, when the triple active long-acting conjugate was administered repeatedly, superior emphysema improvement efficacy was confirmed compared to the excipient control group and the group administered with roflumilast, which is known as a treatment for COPD (* ~ ** p <0.05 ~ 0.01) (Fig. 2).

Through this, it was confirmed that the triple activator long-acting conjugate of the present invention exhibits an effect of improving inflammation and emphysema in lung tissue, thereby having excellent therapeutic efficacy for chronic obstructive pulmonary disease.

Experimental Example 4: Triple activator long-acting conjugate Confirm the effect of improving lung fibrosis

In addition to the effect of improving inflammation in the lung tissue confirmed in the above example, it was attempted to confirm whether the triple activator long-acting conjugate according to the present invention also has an effect of improving lung fibrosis. Therefore, the effect on the differentiation of lung fibroblasts into myofibroblasts and the epithelial mesenchymal transition (EMT) of alveolar epithelial cells, which are known to be important in the lung fibrosis process, was confirmed. For this purpose, the MRC5 cell line, which is a lung fibroblast, and the A549 cell line, which is an alveolar epithelial cell, were used.

Experimental Example 4-1: Confirmation of the inhibitory effect on myofibroblast differentiation of lung fibroblasts

First, MRC5 cells, which are lung fibroblasts, were divided into a vehicle-treated group and a triple activator long-acting conjugate-treated group, and TGF-β1 was co-treated to induce differentiation into myofibroblasts. As a negative control group, a group that was not treated with both TGF-β1 and the triple activator long-acting conjugate was used.

After 48 to 72 hours of co-treatment, the expression levels of α-SMA, collagen1α1, and fibronectin, which are markers of myofibroblast differentiation in each group, were confirmed through quantitative PCR. Statistical treatment evaluated the efficacy of triple activator long-acting conjugates using one-way ANOVA. (* ~ ** p < 0 .05 ~ 0.01)

As a result, it was confirmed that when the triple activator long-acting conjugate was simultaneously treated with TGF-β1, the expression of the myofibroblast differentiation marker decreased compared to the excipient-treated group ( FIG. 3 ).

Experimental Example 4-2: Effect of alveolar epithelial cells to inhibit epithelial-mesenchymal transition (EMT) confirmation

Next, A549 cells were divided into an excipient-treated group and a triple-activator long-acting conjugate-treated group and pre-treated, followed by sequential treatment with TGF-β1 and LPS to induce EMT. As a negative control group, a group not treated with the triple activator long-acting conjugate, TGF-β1, and LPS was used.

After 48 hours, the expression levels of collagen1α1 and collagen3α1, which are EMT markers, in each group were confirmed by quantitative PCR. Statistical treatment evaluated the efficacy of triple activator long-acting conjugates using one-way ANOVA. (* ~ *** p < 0 .05 ~ 0.001)

As a result, it was confirmed that the expression of EMT markers in alveolar epithelial cells was significantly reduced compared to the excipient-treated group when the triple activator long-acting conjugate was pre-treated ( FIG. 4 ).

Experimental Example 4-3: Improvement of fibrosis in vivo check the effect

To confirm in vivo the efficacy of the triple activator long-acting conjugate prepared in the above Examples for improving pulmonary fibrosis, particularly idiopathic pulmonary fibrosis (IPF), which is a representative pulmonary fibrosis, bleomycin (BLM) mice were used. When bleomycin is administered intratracheally, it is known that an epithelial-mesenchymal transition occurs by damaging the DNA of alveolar epithelial cells, and idiopathic fibrosis is induced. In addition, it is known that when the BLM treatment dose is increased above a certain level, the survival rate of the mouse is reduced due to severe lung damage due to idiopathic fibrosis.

Accordingly, the effect of improving lung fibrosis of the triple-activator long-acting conjugate in mice treated with BLM was confirmed, and further, it was confirmed whether the survival rate could be increased by improving lung fibrosis.

IPF mice treated with BLM 1.5U/head in C57BL/6 mice (ORIENT Bio, Busan, Korea) were treated with a vehicle control group, a triple activator long-acting conjugate (3.9 nmol/kg, Q2D, subcutaneous) group, and pirfenidone (pirfenidone). ; 300 mg/kg, QD, oral) administration group, and ambroxol (45 mg/kg, BID, intraperitoneal) administration group, and repeated administration was performed for 2 weeks. After repeated administration for 2 weeks, lung tissue of each mouse was taken as an autopsy, and the degree of fibrosis in the lung tissue was evaluated by masson trichrome staining. Statistical treatment evaluated the efficacy of triple-activator long-acting conjugates using one-way ANOVA (* ~ *** p <0.05 ~ 0.001)

As a result, when repeated administration of the triple activator long-acting conjugate was compared to the excipient control group and the group administered with pirfenidone, which is known as an IPF treatment, it was possible to confirm an excellent effect on reducing the positive area of masson trichrome staining in lung tissue (* ~ *** p < 0.05 to 0.001). In addition, in the case of ambroxol, which is known as an antitussive expectorant remover, it was confirmed that it showed the same level of efficacy as the long-acting conjugate of the triple activator (FIG. 5)

In addition, IPF mice treated with BLM 3.0U/head to C57BL/6 mice (ORIENT Bio, Busan, Korea) were treated with a vehicle control group, a triple activator long-acting conjugate (3.9 nmol/kg, Q2D, subcutaneous) administration group, pirfeni It was divided into a don (300 mg/kg, QD, oral) administration group and amboroxol (45 mg/kg, BID, intraperitoneal) administration group, and repeated administration was performed for 2 weeks, and the survival rate during repeated administration was confirmed.

As a result, it was confirmed that, when the long-acting conjugate of the triple activator was repeatedly administered, the excipient control group, the pirfenidone-administered group, and the ambroxol-administered group showed superior survival rates (FIG. 6).

Through this, it was confirmed that the long-acting conjugate of the triple activator had a therapeutic effect on pulmonary fibrosis or idiopathic pulmonary fibrosis (IPF), thereby improving the survival rate.

It was confirmed that the triple activator long-acting conjugate according to the present invention can improve lung fibrosis by inhibiting myofibroblast differentiation of pulmonary fibroblasts and EMT of alveolar epithelial cells, and also in vivo through the effect of improving lung fibrosis. It was confirmed that fibrosis, particularly idiopathic pulmonary fibrosis, could be effective in preventing and treating.

Taken together, the above results suggest that the triple activator conjugates of the present invention can effectively prevent or treat related lung diseases through improvement of lung inflammation and pulmonary fibrosis, meaning that they can be provided as new therapeutic agents for lung diseases. do.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the claims described below and their equivalents.

<110> HANMI PHARM. CO., LTD. <120> Long acting conjugate of Trigonal glucagon/GLP-1/GIP receptor agonist for use in the treatment of pulmonary disease <130> KPA200028-EN-P1 <150> KR 10-2020-0004379 <151> 2020-01-13 <150> KR 10-2020-0134344 <151> 2020-10-16 <160> 139 <170> KoPatentIn 3.0 <210> 1 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 1 His Xaa Gln Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Asp Gly 1 5 10 15 Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Cys 20 25 30 <210> 2 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 2 His Xaa Gln Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Asp Gly 1 5 10 15 Gln Ala Gln Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Cys 20 25 30 <210> 3 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 3 His Xaa Gln Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Leu Gly 1 5 10 15 Gln Ala Ala Lys Gln Phe Ile Ala Trp Leu Val Lys Gly Gly Gly Pro 20 25 30 Ser Ser Gly Ala Pro Pro Pro Ser Cys 35 40 <210> 4 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 4 His Xaa Gln Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Leu Gly 1 5 10 15 Gln Gln Gln Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Cys 20 25 30 <210> 5 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 5 His Xaa Gln Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Leu Gly 1 5 10 15 Gln Gln Gln Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Gly Gly Pro 20 25 30 Ser Ser Gly Ala Pro Pro Pro Ser Cys 35 40 <210> 6 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 6 His Xaa Gln Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Asp Gly 1 5 10 15 Gln Ala Ala Lys Glu Phe Val Ala Trp Leu Leu Lys Gly Cys 20 25 30 <210> 7 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 7 His Xaa Gln Gly Thr Phe Thr Ser Asp Val Ser Lys Tyr Leu Asp Gly 1 5 10 15 Gln Ala Ala Lys Glu Phe Val Ala Trp Leu Leu Lys Gly Cys 20 25 30 <210> 8 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 8 His Xaa Gln Gly Thr Phe Thr Ser Asp Val Ser Lys Tyr Leu Asp Gly 1 5 10 15 Gln Ala Ala Gln Glu Phe Val Ala Trp Leu Leu Lys Gly Cys 20 25 30 <210> 9 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 9 His Xaa Gln Gly Thr Phe Thr Ser Asp Val Ser Lys Tyr Leu Asp Gly 1 5 10 15 Gln Ala Ala Gln Glu Phe Val Ala Trp Leu Leu Ala Gly Cys 20 25 30 <210> 10 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 10 His Xaa Gln Gly Thr Phe Thr Ser Asp Val Ser Lys Tyr Leu Asp Gly 1 5 10 15 Gln Ala Ala Gln Glu Phe Val Ala Trp Leu Leu Ala Gly Gly Gly Pro 20 25 30 Ser Ser Gly Ala Pro Pro Pro Ser Cys 35 40 <210> 11 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <400> 11 Xaa Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Tyr Leu Asp Ser 1 5 10 15 Arg Arg Gln Gln Leu Phe Val Gln Trp Leu Lys Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser His Gly 35 40 <210> 12 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <400> 12 Xaa Gly Glu Gly Thr Phe Ile Ser Asp Leu Ser Lys Tyr Met Asp Glu 1 5 10 15 Gln Ala Val Gln Leu Phe Val Glu Trp Leu Met Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser His Gly 35 40 <210> 13 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <400> 13 Xaa Gly Glu Gly Thr Phe Ile Ser Asp Tyr Ser Ile Gln Leu Asp Glu 1 5 10 15 Ile Ala Val Gln Asp Phe Val Glu Trp Leu Leu Ala Gln Lys Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser His Gly 35 40 <210> 14 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <400> 14 Xaa Gly Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Gln Leu Asp Glu 1 5 10 15 Ile Ala Val Arg Asp Phe Val Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser His Gly 35 40 <210> 15 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <400> 15 Xaa Gly Gln Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Asp Glu 1 5 10 15 Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser His Gly 35 40 <210> 16 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <400> 16 Xaa Gly Gln Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Asp Ser 1 5 10 15 Glu Ala Gln Gln Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser His Gly 35 40 <210> 17 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <400> 17 Xaa Gly Gln Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Asp Glu 1 5 10 15 Glu Arg Ala Arg Glu Phe Ile Glu Trp Leu Leu Ala Gln Lys Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser His Gly 35 40 <210> 18 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <400> 18 Xaa Gly Gln Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Asp Ser 1 5 10 15 Glu Arg Ala Arg Glu Phe Ile Glu Trp Leu Lys Asn Thr Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser His Gly 35 40 <210> 19 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <400> 19 Xaa Gly Gln Gly Thr Phe Thr Ser Asp Leu Ser Ile Gln Tyr Asp Ser 1 5 10 15 Glu His Gln Arg Asp Phe Ile Glu Trp Leu Lys Asp Thr Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser His Gly 35 40 <210> 20 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <400> 20 Xaa Gly Gln Gly Thr Phe Thr Ser Asp Leu Ser Ile Gln Tyr Glu Glu 1 5 10 15 Glu Ala Gln Gln Asp Phe Val Glu Trp Leu Lys Asp Thr Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser His Gly 35 40 <210> 21 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 21 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Cys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 22 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 22 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Cys Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 23 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 23 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Cys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Ala Gln Lys Gly Lys 20 25 30 Lys Asn Asp Trp Lys His Asn Ile Thr 35 40 <210> 24 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 24 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Cys Arg Ala Lys Glu Phe Val Gln Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 25 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 25 His Xaa Gln Gly Thr Phe Thr Ser Asp Cys Ser Lys Tyr Leu Asp Glu 1 5 10 15 Arg Ala Ala Gln Asp Phe Val Gln Trp Leu Leu Asp Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 26 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 26 His Xaa Gln Gly Thr Phe Thr Ser Asp Cys Ser Lys Tyr Leu Asp Ser 1 5 10 15 Arg Ala Ala Gln Asp Phe Val Gln Trp Leu Leu Asp Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 27 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 27 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Arg Ala Cys Gln Asp Phe Val Gln Trp Leu Leu Asp Gln Gly Gly Pro 20 25 30 Ser Ser Gly Ala Pro Pro Pro Ser 35 40 <210> 28 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 28 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Ala Gln Glu Phe Val Cys Trp Leu Leu Ala Gln Lys Gly Lys 20 25 30 Lys Asn Asp Trp Lys His Asn Ile Thr 35 40 <210> 29 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 29 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Ala Ala Lys Glu Phe Val Gln Trp Leu Leu Asn Thr Cys 20 25 30 <210> 30 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 30 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Ala Gln Lys Glu Phe Val Gln Trp Leu Leu Asp Thr Cys 20 25 30 <210> 31 <211> 29 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 31 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Ala Cys Lys Glu Phe Val Gln Trp Leu Leu Ala Gln 20 25 <210> 32 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 32 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Ala Cys Lys Asp Phe Val Gln Trp Leu Leu Asp Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 33 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 33 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Ala Gln Lys Cys 20 25 30 <210> 34 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 34 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Asn Trp Leu Leu Ala Gln Lys Cys 20 25 30 <210> 35 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 35 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Asn Thr Lys Cys 20 25 30 <210> 36 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 36 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Cys Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Gln Trp Leu Leu Asn Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser Gly 35 40 <210> 37 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 37 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Cys Arg Gln Lys Glu Phe Val Gln Trp Leu Leu Asn Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser Gly 35 40 <210> 38 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 38 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Lys Ser Ser Tyr Leu Asp Glu 1 5 10 15 Arg Ala Ala Gln Asp Phe Val Gln Trp Leu Leu Asp Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser Ser Ser 35 40 <210> 39 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 39 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Gly 1 5 10 15 Gln His Ala Gln Cys Phe Val Ala Trp Leu Leu Ala Gly Gly Gly Pro 20 25 30 Ser Ser Gly Ala Pro Pro Pro Ser 35 40 <210> 40 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 40 His Xaa Gln Gly Thr Phe Thr Ser Asp Lys Ser Lys Tyr Leu Asp Glu 1 5 10 15 Arg Ala Cys Gln Asp Phe Val Gln Trp Leu Leu Asp Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 41 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 41 His Xaa Gln Gly Thr Phe Thr Ser Asp Lys Ser Lys Tyr Leu Asp Glu 1 5 10 15 Cys Ala Ala Gln Asp Phe Val Gln Trp Leu Leu Asp Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 42 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 42 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser Cys 35 40 <210> 43 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 43 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Cys Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 44 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <400> 44 His Gly Gln Gly Thr Phe Thr Ser Asp Cys Ser Lys Gln Leu Asp Gly 1 5 10 15 Gln Ala Ala Gln Glu Phe Val Ala Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 45 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <400> 45 His Gly Gln Gly Thr Phe Thr Ser Asp Cys Ser Lys Tyr Met Asp Gly 1 5 10 15 Gln Ala Ala Gln Asp Phe Val Ala Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 46 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <400> 46 His Gly Gln Gly Thr Phe Thr Ser Asp Cys Ser Lys Tyr Leu Asp Glu 1 5 10 15 Gln His Ala Gln Glu Phe Val Ala Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 47 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <400> 47 His Gly Gln Gly Thr Phe Thr Ser Asp Cys Ser Lys Tyr Leu Asp Gly 1 5 10 15 Gln Arg Ala Gln Glu Phe Val Ala Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 48 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <400> 48 His Gly Gln Gly Thr Phe Thr Ser Asp Cys Ser Lys Tyr Leu Asp Gly 1 5 10 15 Gln Arg Ala Gln Asp Phe Val Asn Trp Leu Leu Ala Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 49 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 49 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Cys Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Asn Thr Lys 20 25 30 <210> 50 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 50 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser Cys 35 40 <210> 51 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 51 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Gln Trp Leu Leu Asn Thr Cys 20 25 30 <210> 52 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 52 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Gln Trp Leu Leu Asp Thr Cys 20 25 30 <210> 53 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 53 His Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Ala Gln Cys 20 25 30 <210> 54 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 54 His Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asp Trp Leu Leu Ala Glu Cys 20 25 30 <210> 55 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 55 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Ala Gln Cys 20 25 30 <210> 56 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 56 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Asn Trp Leu Leu Ala Gln Cys 20 25 30 <210> 57 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 57 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Asn Thr Cys 20 25 30 <210> 58 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 58 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Gln Trp Leu Leu Asn Thr Lys Cys 20 25 30 <210> 59 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 59 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Cys Met Asp Glu 1 5 10 15 Lys His Gln Lys Asp Phe Val Asn Trp Leu Leu Asn Thr Lys 20 25 30 <210> 60 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 60 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Lys His Cys Lys Asp Phe Val Asn Trp Leu Leu Asn Thr Lys 20 25 30 <210> 61 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 61 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile Ala Cys Lys Asp Phe Val Asn Trp Leu Leu Asn Thr Lys 20 25 30 <210> 62 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 62 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Lys Ser Lys Tyr Leu Asp Glu 1 5 10 15 Arg Ala Ala Gln Asp Phe Val Gln Trp Leu Leu Asp Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 63 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <400> 63 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Cys Ser Lys Tyr Leu Asp Glu 1 5 10 15 Arg Ala Ala Gln Asp Phe Val Gln Trp Leu Leu Asp Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 64 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 64 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Cys Ala Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 65 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 65 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Cys Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 66 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 66 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Cys Arg Ala Lys Asp Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 67 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 67 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Cys Ala Ala Lys Asp Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 68 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 68 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Cys Leu Asp Glu 1 5 10 15 Lys Ala Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 69 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 69 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Cys Leu Asp Glu 1 5 10 15 Arg Ala Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 70 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 70 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Cys Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Asp Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 71 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 71 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Arg Ala Cys Lys Asp Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 72 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 72 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Cys Ser Lys Tyr Leu Asp Glu 1 5 10 15 Arg Ala Ala Lys Asp Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 73 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 73 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Cys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 74 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 74 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Cys Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 75 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 75 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Ala Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser Cys 35 40 <210> 76 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 76 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Asp Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser Cys 35 40 <210> 77 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 77 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Ala Ala Lys Asp Phe Val Gln Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser Cys 35 40 <210> 78 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 78 His Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Gln Trp Leu Leu Asp Thr Lys Cys 20 25 30 <210> 79 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 79 His Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Ala Gln Lys Cys 20 25 30 <210> 80 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 80 His Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asp Trp Leu Leu Ala Glu Lys Cys 20 25 30 <210> 81 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 81 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Gln Trp Leu Leu Asn Thr Cys 20 25 30 <210> 82 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 82 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Gln Trp Leu Leu Asp Thr Cys 20 25 30 <210> 83 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 83 Xaa Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Ala Gln Cys 20 25 30 <210> 84 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 84 Xaa Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asp Trp Leu Leu Ala Glu Cys 20 25 30 <210> 85 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 85 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Ala Gln Cys 20 25 30 <210> 86 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 86 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Asn Trp Leu Leu Ala Gln Cys 20 25 30 <210> 87 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 87 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Asn Thr Cys 20 25 30 <210> 88 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 88 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Gln Trp Leu Leu Asn Thr Lys Cys 20 25 30 <210> 89 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 89 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Gln Trp Leu Leu Asp Thr Lys Cys 20 25 30 <210> 90 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 90 Xaa Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Ala Gln Lys Cys 20 25 30 <210> 91 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 91 Xaa Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asp Trp Leu Leu Ala Glu Lys Cys 20 25 30 <210> 92 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 92 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Ala Gln Lys Cys 20 25 30 <210> 93 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 93 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Gln Lys Glu Phe Val Asn Trp Leu Leu Ala Gln Lys Cys 20 25 30 <210> 94 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (1) <223> Xaa is 4-imidazoacetyl (CA) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 94 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ala Met Asp Glu 1 5 10 15 Ile His Gln Lys Asp Phe Val Asn Trp Leu Leu Asn Thr Lys Cys 20 25 30 <210> 95 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 95 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Cys His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 96 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 96 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Asp His Cys Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 97 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 97 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 1 5 10 15 Lys Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Asp Cys His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser 35 <210> 98 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 98 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Ala Leu Asp Glu 1 5 10 15 Lys Ala Ala Lys Glu Phe Val Asn Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser Cys 35 40 <210> 99 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 99 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Ala Leu Asp Glu 1 5 10 15 Lys Ala Ala Lys Asp Phe Val Asn Trp Leu Leu Asp His His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser Cys 35 40 <210> 100 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 100 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Ala Leu Asp Glu 1 5 10 15 Lys Ala Ala Lys Glu Phe Val Gln Trp Leu Leu Asp Gln His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser Cys 35 40 <210> 101 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 101 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Ala Leu Asp Glu 1 5 10 15 Lys Ala Ala Lys Glu Phe Val Asn Trp Leu Leu Asp Gln His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser Cys 35 40 <210> 102 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> Trigonal agonist <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 2-aminoisobutyric acid (Aib) <220> <221> MISC_FEATURE <222> (16)..(20) <223> amino acids at positions 16 and 20 form a ring <400> 102 Tyr Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Ala Leu Asp Glu 1 5 10 15 Lys Ala Ala Lys Asp Phe Val Asn Trp Leu Leu Asp Gln His Pro Ser 20 25 30 Ser Gly Gln Pro Pro Pro Ser Cys 35 40 <210> 103 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> General Formula 1 <220> <221> MISC_FEATURE <222> (1) <223> Xaa is His (H), 4-imidazoacetyl (CA), or Tyr (Y) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is Gly (G), alpha-methyl-glutamic acid, or Aib (2-aminoisobutyric acid) <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Glu (E) or Gln (Q) <220> <221> MISC_FEATURE <222> (7) <223> Xaa is Thr (T) or Ile (I) <220> <221> MISC_FEATURE <222> (10) <223> Xaa is Leu (L), Tyr (Y), Lys (K), Cys (C), or Val (V) <220> <221> MISC_FEATURE <222> (12) <223> Xaa is Lys (K), Ser (S), or Ile (I) <220> <221> MISC_FEATURE <222> (13) <223> Xaa is Gln (Q), Tyr (Y), Ala (A), or Cys (C) <220> <221> MISC_FEATURE <222> (14) <223> Xaa is Leu (L), Met (M), or Tyr (Y) <220> <221> MISC_FEATURE <222> (15) <223> Xaa is Cys (C), Asp (D), Glu (E), or Leu (L) <220> <221> MISC_FEATURE <222> (16) <223> Xaa is Gly (G), Glu (E), or Ser (S) <220> <221> MISC_FEATURE <222> (17) <223> Xaa is Gln (Q), Arg (R), Ile (I), Glu (E), Cys (C), or Lys (K) <220> <221> MISC_FEATURE <222> (18) <223> Xaa is Ala (A), Gln (Q), Arg (R), or His (H) <220> <221> MISC_FEATURE <222> (19) <223> Xaa is Ala (A), Gln (Q), Cys (C), or Val (V) <220> <221> MISC_FEATURE <222> (20) <223> Xaa is Lys (K), Gln (Q), or Arg (R) <220> <221> MISC_FEATURE <222> (21) <223> Xaa is Glu (E), Gln (Q), Leu (L), Cys (C), or Asp (D) <220> <221> MISC_FEATURE <222> (23) <223> Xaa is Ile (I) or Val (V) <220> <221> MISC_FEATURE <222> (24) <223> Xaa is Ala (A), Gln (Q), Cys (C), Asn (N), Asp (D), or Glu (E) <220> <221> MISC_FEATURE <222> (27) <223> Xaa is Val (V), Leu (L), Lys (K), or Met (M) <220> <221> MISC_FEATURE <222> (28) <223> Xaa is Cys (C), Lys (K), Ala (A), Asn (N), or Asp (D) <220> <221> MISC_FEATURE <222> (29) <223> Xaa is Cys (C), Gly (G), Gln (Q), Thr (T), Glu (E), or His (H) <220> <221> MISC_FEATURE <222> (30) <223> Xaa is Cys (C), Gly (G), Lys (K), or His (H), or is absent; and Xaa can be further linked to R1, wherein R1 is Cys (C), GKKNDWKHNIT, m-SSGAPPPS-n, or m-SSGQPPPS-n, or is absent, and wherein m is -Cys-, -Pro-, or -Gly-Pro-, and n is -Cys-, -Gly-, -Ser-, or -His-Gly-, or is absent <400> 103 Xaa Xaa Xaa Gly Thr Phe Xaa Ser Asp Xaa Ser Xaa Xaa Xaa Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Phe Xaa Xaa Trp Leu Xaa Xaa Xaa Xaa 20 25 30 <210> 104 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> General Formula 2 <220> <221> MISC_FEATURE <222> (1) <223> Xaa is His (H), 4-imidazoacetyl (CA), or Tyr (Y) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is Gly (G), alpha-methyl-glutamic acid, or Aib (2-aminoisobutyric acid) <220> <221> MISC_FEATURE <222> (10) <223> Xaa is Tyr (Y) or Cys (C) <220> <221> MISC_FEATURE <222> (13) <223> Xaa is Gln (Q), Tyr (Y), Ala (A), or Cys (C) <220> <221> MISC_FEATURE <222> (14) <223> Xaa is Leu (L), Met (M), or Tyr (Y) <220> <221> MISC_FEATURE <222> (15) <223> Xaa is Asp (D), Glu (E), or Leu (L) <220> <221> MISC_FEATURE <222> (16) <223> Xaa is Gly (G), Glu (E), or Ser (S) <220> <221> MISC_FEATURE <222> (17) <223> Xaa is Gln (Q), Arg (R), Ile (I), Glu (E), Cys (C), or Lys (K) <220> <221> MISC_FEATURE <222> (18) <223> Xaa is Ala (A), Gln (Q), Arg (R), or His (H) <220> <221> MISC_FEATURE <222> (19) <223> Xaa is Ala (A), Gln (Q), Cys (C), or Val (V) <220> <221> MISC_FEATURE <222> (20) <223> Xaa is Lys (K), Gln (Q), or Arg (R) <220> <221> MISC_FEATURE <222> (21) <223> Xaa is Glu (E), Gln (Q), Leu (L), Cys (C), or Asp (D) <220> <221> MISC_FEATURE <222> (23) <223> Xaa is Ile (I) or Val (V) <220> <221> MISC_FEATURE <222> (24) <223> Xaa is Ala (A), Gln (Q), Cys (C), Asn (N), or Glu (E) <220> <221> MISC_FEATURE <222> (28) <223> Xaa is Cys (C), Lys (K), Asn (N), or Asp (D) <220> <221> MISC_FEATURE <222> (29) <223> Xaa is Cys (C), Gly (G), Gln (Q), or His (H) <220> <221> MISC_FEATURE <222> (30) <223> Xaa is Cys (C), Gly (G), Lys (K), or His (H) <220> <221> MISC_FEATURE <222> (31) <223> Xaa is Pro (P) or Cys (C) <220> <221> MISC_FEATURE <222> (40) <223> Xaa is Cys (C), or is absent <400> 104 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Xaa Ser Lys Xaa Xaa Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Phe Xaa Xaa Trp Leu Leu Xaa Xaa Xaa Xaa Ser 20 25 30 Ser Gly Gln Pro Pro Ser Xaa 35 40 <210> 105 <211> 40 <212> PRT <213> Artificial Sequence <220> <223> General Formula 3 <220> <221> MISC_FEATURE <222> (1) <223> Xaa is His (H) or Tyr (Y) <220> <221> MISC_FEATURE <222> (2) <223> Xaa is alpha-methyl-glutamic acid, or Aib (2-aminoisobutyric acid) <220> <221> MISC_FEATURE <222> (13) <223> Xaa is Tyr (Y), Ala (A), or Cys (C) <220> <221> MISC_FEATURE <222> (17) <223> Xaa is Arg (R), Cys (C), or Lys (K) <220> <221> MISC_FEATURE <222> (18) <223> Xaa is Ala (A) or Arg (R) <220> <221> MISC_FEATURE <222> (19) <223> Xaa is Ala (A) or Cys (C) <220> <221> MISC_FEATURE <222> (21) <223> Xaa is Glu (E) or Asp (D) <220> <221> MISC_FEATURE <222> (24) <223> Xaa is Gln (Q) or Asn (N) <220> <221> MISC_FEATURE <222> (28) <223> Xaa is Cys (C) or Asp (D) <220> <221> MISC_FEATURE <222> (29) <223> Xaa is Cys (C), Gln (Q), or His (H) <220> <221> MISC_FEATURE <222> (30) <223> Xaa is Cys (C) or His (H) <220> <221> MISC_FEATURE <222> (31) <223> Xaa is Pro (P) or Cys (C) <220> <221> MISC_FEATURE <222> (40) <223> Xaa is Cys (C), or is absent <400> 105 Xaa Xaa Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Xaa Leu Asp Glu 1 5 10 15 Xaa Xaa Xaa Lys Xaa Phe Val Xaa Trp Leu Leu Xaa Xaa Xaa Xaa Ser 20 25 30 Ser Gly Gln Pro Pro Ser Xaa 35 40 <210> 106 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 106 Gly Lys Lys Asn Asp Trp Lys His Asn Ile Thr 1 5 10 <210> 107 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 107 Ser Ser Gly Ala Pro Pro Pro Ser 1 5 <210> 108 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 108 Ser Ser Gly Gln Pro Pro Pro Ser 1 5 <210> 109 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 109 Cys Ser Ser Gly Gln Pro Pro Pro Ser 1 5 <210> 110 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 110 Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser 1 5 10 <210> 111 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 111 Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser Cys 1 5 10 <210> 112 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 112 Pro Ser Ser Gly Ala Pro Pro Pro Ser 1 5 <210> 113 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 113 Pro Ser Ser Gly Ala Pro Pro Pro Ser Gly 1 5 10 <210> 114 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 114 Pro Ser Ser Gly Ala Pro Pro Pro Ser His Gly 1 5 10 <210> 115 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 115 Pro Ser Ser Gly Ala Pro Pro Pro Ser Ser 1 5 10 <210> 116 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 116 Pro Ser Ser Gly Gln Pro Pro Pro Ser 1 5 <210> 117 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> R1 <400> 117 Pro Ser Ser Gly Gln Pro Pro Pro Ser Cys 1 5 10 <210> 118 <211> 29 <212> PRT <213> Homo sapiens <400> 118 His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser 1 5 10 15 Arg Arg Ala Gln Asp Phe Val Gln Trp Leu Met Asn Thr 20 25 <210> 119 <211> 12 <212> PRT <213> HOMO SAPIENS <400> 119 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro 1 5 10 <210> 120 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 120 Glu Ser Lys Tyr Gly Pro Pro Ser Cys Pro 1 5 10 <210> 121 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 121 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Pro 1 5 10 <210> 122 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 122 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser 1 5 10 <210> 123 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 123 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro 1 5 10 <210> 124 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 124 Lys Tyr Gly Pro Pro Cys Pro Ser 1 5 <210> 125 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 125 Glu Ser Lys Tyr Gly Pro Pro Cys 1 5 <210> 126 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 126 Glu Lys Tyr Gly Pro Pro Cys 1 5 <210> 127 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 127 Glu Ser Pro Ser Cys Pro 1 5 <210> 128 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 128 Glu Pro Ser Cys Pro 1 5 <210> 129 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 129 Pro Ser Cys Pro One <210> 130 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 130 Glu Ser Lys Tyr Gly Pro Ser Cys Pro 1 5 10 <210> 131 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 131 Lys Tyr Gly Pro Pro Ser Cys Pro 1 5 <210> 132 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 132 Glu Ser Lys Tyr Gly Pro Ser Cys Pro 1 5 <210> 133 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 133 Glu Ser Lys Tyr Gly Pro Pro Cys 1 5 <210> 134 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 134 Lys Tyr Gly Pro Pro Cys Pro 1 5 <210> 135 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 135 Glu Ser Lys Pro Ser Cys Pro 1 5 <210> 136 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 136 Glu Ser Pro Ser Cys Pro 1 5 <210> 137 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 137 Glu Pro Ser Cys One <210> 138 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Variant of hinge region <400> 138 Ser Cys Pro One <210> 139 <211> 221 <212> PRT <213> homo sapiens <400> 139 Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu 1 5 10 15 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 20 25 30 Val Thr Cys Val Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln 35 40 45 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 50 55 60 Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu 65 70 75 80 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 85 90 95 Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys 100 105 110 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 115 120 125 Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 130 135 140 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 145 150 155 160 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 165 170 175 Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln 180 185 190 Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 195 200 205 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 210 215 220

Claims (19)

As a pharmaceutical composition for the prevention or treatment of lung disease,
pharmaceutically acceptable excipients and
A pharmaceutical composition comprising a pharmaceutically effective amount of a peptide comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 102.
The pharmaceutical composition according to claim 1, wherein the peptide is in the form of a long-acting conjugate, and the long-acting conjugate is represented by the following formula (1):
[Formula 1]
X - L - F
With the proviso that X is a peptide of any one of SEQ ID NOs: 1 to 102;
L is a linker containing an ethylene glycol repeating unit,
F is an immunoglobulin Fc region,
- represents a covalent linkage between X and L and between L and F.
The pharmaceutical composition according to claim 1 or 2, wherein the peptide is amidated at its C-terminus.
The method according to claim 1 or 2, wherein the peptide is selected from the group consisting of SEQ ID NOs: 21, 22, 42, 43, 50, 64, 66, 67, 70, 71, 76, 77, 96, 97 and 100. A pharmaceutical composition comprising an amino acid sequence.
The pharmaceutical composition according to claim 4, wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 21, 22, 42, 43, 50, 77 and 96.
The pharmaceutical composition according to claim 1 or 2, wherein the peptide sequence forms a ring between amino acids 16 and 20 from the N-terminus.
The pharmaceutical composition according to claim 2, wherein the formula weight of the ethylene glycol repeating unit moiety in L is in the range of 1 to 100 kDa.
The pharmaceutical composition according to claim 2, wherein F is an IgG Fc region.
The method of claim 1 or 2, wherein the lung disease is interstitial lung disease (ILD), progressive fibrosing interstitial lung disease (PF-ILD), idiopathic interstitial pneumonia (idiopathic interstitial) pneumonias (IIP), non-specific interstitial pneumonia (NSIP), pulmonary fibrosis, fibrosing interstitial lung diseases (FILD), idiopathic pulmonary fibrosis (IPF) , alveolitis, pneumonia, emphysema, bronchitis, chronic obstructive pulmonary disease, combined pulmonary fibrosis and emphysema (CPFE), asthma ( asthma), or a respiratory infectious disease, a pharmaceutical composition.
The pharmaceutical composition of claim 9, wherein the respiratory infectious disease is a respiratory viral, bacterial, mycoplasma, or fungal infection.
11. The method of claim 10, wherein the respiratory virus is adenovirus, vaccinia virus, herpes simplex virus, parainfluenza virus, rhinovirus, varicella virus ( varicella Zoster Virus, measle virus, respiratorysyncytial virus, Dengue virus, human immunodeficiency virus (HIV), influenza virus, coronavirus, severe acute respiratory syndrome virus ( Severe acute respiratory syndrome associated virus; SARS-associated virus), and any one selected from the group consisting of middle east respiratory syndrome coronavirus (MERS-CoV), a pharmaceutical composition.
The pharmaceutical composition of claim 11 , wherein the coronavirus is SARS-CoV-2.
According to claim 1 or 2, wherein the pharmaceutical composition is administered (i) macrophage (macrophage) activity inhibition, or (ii) the expression of IL-1β, IL-6, IL-12, or TNF-α when administered To reduce the, pharmaceutical composition.
The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition has one or more of the following properties upon administration:
(i) inhibition of myofibroblast differentiation;
(ii) decreased expression of α-SMA, collagen1α1, or fibronectin;
(iii) inhibition of epithelial mesenchymal transition (EMT) of alveolar epithelial cells; and
(iv) decreased expression of collagen1α1, or collagen1α3.
The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is additionally administered with mucolytic agents or a pharmaceutically acceptable salt thereof.
16. The method of claim 15, wherein the mucolytic agent is ambroxol, N-acetylcysteine (N-acetylcystein), N-acetylin (N-acetylin), carbocysteine (carbocysteine), domiodol (domiodol), fudosteine, bromhexine, erdosteine, letostine, lysozyme, mesna, sobrerol, stepronin, thio Pronin, tyloxapol, carbocisteine, dornase alfa, eprazinone, letosteine, neltenexine, and mecysteine ) at least one selected from the group consisting of, a pharmaceutical composition.
The pharmaceutical composition according to claim 14, wherein the peptide and the mucolytic agent or a pharmaceutically acceptable salt thereof are administered simultaneously, sequentially, or in reverse order.
The pharmaceutical composition of claim 9, wherein the lung disease is pneumonia (pulmonary inflammation) or pulmonary fibrosis caused by coronavirus infection-19 (COVID-19).
The pharmaceutical composition according to claim 2, wherein the region F is a dimer consisting of two polypeptide chains, and one end of L is linked to only one of the two polypeptide chains.

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